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Surface Modification Studies On Sulphide Minerals Using Bioreagents
Surface Modification Studies On Sulphide Minerals Using Bioreagents
Abstract
The interaction of galena and sphalerite minerals with the metabolite obtained from Bacillus polymyxa has been examined
through adsorption, electrokinetic, microflotation and flocculation tests. The adsorption density of the carbohydrate
component of the metabolite for sphalerite exhibits a characteristic maximum in the pH range of 6 – 7, while in the case of
galena the amount adsorbed increases with increase of pH. On the other hand, the adsorption density of the bacterial protein
shows a continuous decrease with increase of pH, for both the minerals. The adsorption affinity of both the metabolic
components is higher for galena vis-à-vis sphalerite. The electrophoretic mobility of the chosen minerals becomes less
negative after interaction with the metabolite, in proportion with the time of interaction. Interestingly, the isoelectric point of
sphalerite is shifted to less acidic values after treatment with the metabolite, but that of galena is unaltered. Bioflotation and
bioflocculation studies on a synthetic mixture of galena and sphalerite demonstrate that galena can be selectively depressed or
flocculated from sphalerite under appropriate conditions. Co-precipitation tests confirm complexation of lead and zinc species
with the metabolic products, in the bulk solution. Possible mechanisms of interaction between the chosen sulphide minerals
and the bioreagents are discussed.
D 2003 Elsevier B.V. All rights reserved.
Keywords: galena; sphalerite; Bacillus polymyxa metabolite; carbohydrate; protein; bioflotation and bioflocculation
0301-7516/$ - see front matter D 2003 Elsevier B.V. All rights reserved.
doi:10.1016/S0301-7516(03)00097-8
176 S. Subramanian et al. / Int. J. Miner. Process. 72 (2003) 175–188
mineral beneficiation and coal preparation, include sphalerite and galena were respectively found to be
their utilisation as flocculants (Schneider et al., 1994; 1.43 and 0.99 m2/g.
Dubel et al., 1992; Attia, 1987), flotation collectors The strain of B. polymyxa (NCIM 2539) used in
(Smith et al., 1993) and flotation modifiers (Elzeky this study was obtained from the National Collection
and Attia, 1987; Atkins et al., 1987; Rao et al., 1992; of Industrial Microorganisms (NCIM), National
Zheng et al., 2001). Detailed biobeneficiation studies Chemical Laboratory, Pune. The bacteria were sub-
carried out using Bacillus polymyxa with respect to cultured in the laboratory using the modified Brom-
iron ore and bauxite processing have been reported field medium as per the procedure previously
(Deo and Natarajan, 1997, 1998; Natarajan and Deo, described (Anand et al., 1996).
2001). The utility of Thiobacillus thiooxidans and The starch sample used in this study for the
B. polymyxa in the selective depression and floccula- calibration of the extracellular polysaccharide fraction
tion of galena from its mixture with sphalerite has been was obtained from BDH Limited, Poole, England.
recently brought out by us (Santhiya et al., 2001a,b,c). Potassium nitrate was used to maintain the ionic
It is well known that both organic and inorganic strength, while nitric acid and potassium hydroxide
reagents like different types of mineral acids, fatty were used as pH modifiers. All the reagents used in
acids, polysaccharides, proteins and chelating agents this study were of analytical grade. De-ionised double
are generated by microorganisms. These metabolic distilled water with a final conductivity of < 1.5
products have advantages over their chemical counter- Amhos was used for all the tests.
parts in biodegradability and effectiveness at extreme
temperatures or pH and in having lower toxicity. 2.1. Separation of metabolite from the bacterial
Motivated by this, the present investigation was taken culture
up to study the surface chemical changes brought
about on galena and sphalerite minerals using the The bacterium B. polymyxa was grown by adding
metabolite secreted by B. polymyxa during its growth. 10% v/v of an active inoculum to the Bromfield
The feasibility of selective flotation and flocculation medium and incubated at 30 jC on a Remi rotary
in the galena –sphalerite system has been examined in shaker at 240 rpm for 48 h. The growth conditions
the presence of the bacterial carbohydrate and protein and the composition of the medium have been kept
components, and the interaction mechanisms have fixed in all experiments. Precautions were taken to
been ascertained. prevent contamination by undesirable microorganisms
by maintaining aseptic conditions. This fully grown
culture was centrifuged at 10,000 rpm for 10 min at
2. Experimental materials and methods 10 jC to separate the cells from the extracellular
materials. The supernatant (metabolite) obtained was
Mineral samples of sphalerite and galena were filtered through a sterile Millipore membrane filter
obtained from Gregory, Bottley and Lloyd, UK. paper of pore size 0.2 Am using a Tarsons membrane
Mineralogical studies as well as X-ray powder dif- filter holder.
fraction data indicated that the samples were of high
purity. The above samples were dry ground using a 2.2. Adsorption studies
porcelain ball mill and then sieved through 105, 63
and 37 Am BSS sieves. The 37 Am size fraction For the adsorption tests, 1 g of the mineral powder
was further ground finer in a Retsch mortar grinder. of mean (d50) size c 5 Am was taken and pulped to
The mean (d50) particle size of this fraction was found 50 ml. This was followed by the addition of metab-
to be c 5 Am using a Malvern Mastersizer model olite of desired concentration and pre-adjusted pH
3000 particle size analyser. This fraction was used for equivalent to that of the suspension pH. The ionic
the adsorption, electrokinetic and flocculation studies. strength was kept at 10 3 M using KNO3 and the
For the microflotation experiments, 105 + 63 Am total volume was made up to 100 ml. The suspension
fraction of galena and sphalerite was used. The BET was agitated for 4 h at 200 rpm in a Remi orbital
nitrogen specific surface areas of c 5 Am fraction of shaking incubator at 27 F 2 jC. Subsequently, the
S. Subramanian et al. / Int. J. Miner. Process. 72 (2003) 175–188 177
suspension was centrifuged at 5000 rpm using a 2.5. Differential flotation tests
Sorvall RC-5B refrigerated high speed centrifuge
for 10 min and filtered through a Whatman 42 filter In these experiments, 0.5 g each of galena and
paper. The clear supernatant solution was analysed sphalerite (1:1) mixture, of particle size ( 105 + 63)
for the residual carbohydrate and protein concentra- Am was interacted with the metabolite at its natural pH
tions using a Shimadzu UV-260 UV-vis spectropho- of 3.2– 3.4 and at pH 9– 9.5 for 15 min prior to
tometer as per standard procedures (Dubois et al., flotation. The same procedure as adopted for the
1956; Bradford, 1976). flotation of the individual minerals was followed.
The lead and zinc contents in the concentrate and
2.3. Electrokinetic measurements tailing fractions were separately analysed using a
Jobin Yvon JY 24 inductively coupled plasma emis-
Electrophoretic mobilities of sphalerite as well as sion spectrometer.
galena were determined in the presence and absence
of metabolite using a Malvern 3000 model Zetasizer. 2.6. Flocculation tests
A known concentration of the metabolite solution at
a desired pH was added slowly to the suspension The flocculation test procedure was standardised as
containing 10 mg of the solids under agitated follows: 0.5 g of galena or sphalerite of c 5Am size
conditions at the same pH and made up to 100 was suspended in a volume of 100 ml of the metab-
ml. In all these experiments, the pH of the suspen- olite of pre-adjusted pH in the range of 3 –11 in a 100-
sion was adjusted to a desired value using AR grade ml measuring jar. The measuring jar was gently
nitric acid or potassium hydroxide and the ionic tumbled 10 times and allowed to stand for 2 min.
strength was maintained at 10 3 M KNO3. The Exactly 90 ml of the supernatant was siphoned out
suspension was allowed to equilibrate for 1 h and into a clean, 100 ml beaker, filtered through a What-
ultrasonicated for 30 s before measuring its electro- man 42 filter paper and the residue was dried and
phoretic mobility. The effects of pH and interaction weighed. The percentage weights of the solids dis-
time on the electrophoretic mobilities of the minerals persed as well as those settled were determined.
were determined.
2.7. Selective flocculation tests
2.4. Microflotation studies
In these experiments, 0.25 g each of galena and
Microflotation tests were carried out using a mod- sphalerite (1:1) of particle size c 5 Am was initially
ified Hallimond tube (Fuerstenau et al., 1957). Potas- mixed and interacted with 100 ml of the metabolite at
sium isopropyl xanthate (10 4M) was used as a pH 3 –3.5 and at pH 9 –9.5 for 2 min in a 100-ml
collector for both sphalerite and galena, while measuring jar. The same procedure as described for
10 6M CuSO4 was used as an activator for sphalerite. the flocculation of the individual minerals was adop-
For the flotation tests, 1 g of ( 105 + 63) Am size ted. The lead and zinc contents in the dispersed and
mineral sample was pulped to a final volume of 200 ml flocculated portions were analysed using a Jobin
with double distilled water after conditioning with the Yvon JY24 inductively coupled plasma emission
chosen reagents at a desired pH for 15 min using a spectrometer.
magnetic stirrer. The sample was then transferred to the
Hallimond tube and nitrogen gas was passed at the rate 2.8. Co-precipitation studies
of 40 ml per minute. After flotation for 3 min, the
concentrate and the tailing fractions were separately In these studies, a known amount of Pb(NO3)2/
filtered, dried and weighted. The discoveries are ex- Zn(NO3)2 solution was mixed with 100 ml of the
pressed on a weight % basis. In experiments where the metabolite in a such a way that the final concentration
metabolite was added, the sample was further condi- of lead or zinc was 50 ppm in 100 ml solution. The pH
tioned for 15 min at a given pH with it, prior to was adjusted to a desired value in the range of 3 –11 by
flotation. adding either nitric acid or potassium hydroxide solu-
178 S. Subramanian et al. / Int. J. Miner. Process. 72 (2003) 175–188
The metabolite obtained from the fully grown adsorbed increases with increase of pH and attains a
culture of B. polymyxa after 48 h of growth was region of higher adsorption density in the pH range
separated from the bacterial cells and interacted with of 6 – 8.5 and thereafter decreases for sphalerite.
the minerals. The adsorption of the protein and Adsorption maxima around pH 7.5 – 8 have been
exopolysaccharide (carbohydrate) components of the observed for the adsorption of polysaccharides such
metabolite was studied as a function of pH and the as dextrin and guar gum onto sphalerite (Rath and
adsorption isotherms were also determined. Subramanian, 1999a,b). The adsorption density of
the bacterial carbohydrate continuously increases
3.1.1. Effect of pH with increase of pH in the case of galena, although
The adsorption density of carbohydrate from B. a distinct maximum is not apparent. Earlier inves-
polymyxa metabolite for sphalerite and galena as a tigations had revealed a maximum around pH 11.5
function of pH is portrayed is Fig. 1. The amount for the adsorption of dextrin and guar gum onto
galena (Liu and Laskowski, 1989a; Rath and Sub-
ramanian, 1999a,b). It is noteworthy that the maxi-
mum adsorption density of carbohydrate for galena
(160 mg/m2) is about 50 times higher than that for
sphalerite (3.2 mg/m2).
The adsorption of protein from B. polymyxa me-
tabolite onto galena and sphalerite as a function of pH
is shown in Fig. 2. The adsorption density decreases
with increase of pH for both galena and sphalerite.
Additionally, the amount of bacterial protein adsorbed
onto galena is slightly higher than that onto sphalerite.
The functional groups present in the protein, their
charge characteristics and conformation govern the
adsorption process. Most proteins have isoelectric
points in the pH range of 3 – 6 and the reduced
adsorption in the alkaline pH range may be attributed
Fig. 1. Adsorption density of carbohydrate from metabolite of B. to electrostatic repulsion between the protein and the
polymyxa for sphalerite and galena as a function of pH. sulphide mineral.
S. Subramanian et al. / Int. J. Miner. Process. 72 (2003) 175–188 179
3.1.2. Adsorption isotherms 7.1– 7.2 is the highest followed by that at pH 3.4 –3.6,
The adsorption isotherms of carbohydrate, from the while the isotherm at pH 11.1– 11.2 has the lowest
metabolite of B. polymyxa, for sphalerite at three adsorption density. The isotherms at pH 7.1 and 11.1
different pH values are depicted in Fig. 3(a). For the resemble the S-2 type of the Giles classification (Giles
isotherm at pH 7.1 –7.2, the adsorption density ini- et al., 1960), while that at pH 3.4 may be categorised
tially increases sharply up to 1000 ppm concentration as L-1 type.
and thereafter tends to attain a plateau level of Fig. 3(b) shows the adsorption isotherms of bacte-
adsorption. At pH 3.4 –3.6, the adsorption density rial carbohydrate for galena at different pH values.
marginally increases with increase in equilibrium The isotherm at pH 11 shows the maximum adsorp-
concentration up to about 3000 ppm followed by a tion density followed by those at pH 7 and 3 in that
sharp increase. The isotherm at pH 11.1 –11.2 indi- order. The isotherm at c pH 11 shows an increase in
cates a very feeble increase in the adsorption density the adsorption density up to about 1600 ppm equilib-
up to about 2000 ppm and thereafter attains a satura- rium concentration, and subsequently tends to attain a
tion value. These results parallel the pH trend ob- plateau level. The isotherms at about pH 7 and 3 show
served earlier (Fig. 1). The adsorption density at pH a similar trend, except that the plateau level of
adsorption is attained around 2000 ppm concentration.
All the isotherms resemble the S-2 type of the Giles
classification (Giles et al., 1960).
A comparison of Fig. 3(a) and (b) unambiguously
reveals that the adsorption density of bacterial carbo-
hydrate for galena is significantly higher compared to
that for sphalerite. This is in agreement with that
observed in Fig. 1.
The adsorption isotherms of protein from B. poly-
myxa metabolite for sphalerite and galena are shown
in Fig. 4(a) and (b), respectively. In the case of
sphalerite, the adsorption density decreases with in-
crease of pH as observed earlier (Fig. 2). From Fig.
4a, it is evident that the adsorption density initially
increases steeply for the isotherms at c pH 3.5 and 7
and thereafter a decrease in the slope of the adsorp-
tion density takes place. The amount of protein
adsorbed at pH 11 is significantly less. The isotherms
at pH 3.5 and 7 may be classified as belonging to the
S-3 type, while those at pH 11 resemble the low
affinity S-2 type of the Giles category (Giles et al.,
1960). With respect to galena (Fig. 4b), the adsorp-
tion density at pH 3.5 is higher than that at pH 7.1
and 11.1. The isotherms at pH 3.5 and 7.1 show a
steep rise in the adsorption density at lower concen-
trations followed by reduced adsorption at higher
concentrations. On the other hand, at pH 11 the
amount adsorbed is substantially less, presumably
due to electrostatic repulsion between the negatively
charged mineral surface and anionic component of
Fig. 3. (a) Adsorption isotherms of carbohydrate from metabolite of the bacterial protein. The isotherms for galena resem-
B. polymyxa for sphalerite. (b) Adsorption isotherms of carbohy- ble those obtained for sphalerite and can be categor-
drate from metabolite of B. polymyxa for galena. ised in a similar manner. The magnitude of protein
180 S. Subramanian et al. / Int. J. Miner. Process. 72 (2003) 175–188
3.5.2. Effect of pH
The effect of pH on the settling behaviour of
sphalerite and galena in the absence and presence of
B. polymyxa metabolite is depicted in Fig. 8. In the
absence of the metabolite, the amount of sphalerite
settled decreases from about 35% at pH 3 to 17% at
pH 11, while the amount of galena settled increases
from 30% at pH 3 to about 55% at pH 11. In the
presence of the metabolite, the percentage sphalerite
settled is about 10% in the pH range investigated. On
the other hand, the percentage galena flocculated
steeply increases from about 67% at pH 3.4 to about
98% at pH 11.2, in the presence of the metabolite. A
careful scrutiny of Fig. 8 indicates that in the presence
of the metabolite, galena can be selectively flocculat-
ed from sphalerite in the pH range of 9 –11 using B.
polymyxa metabolite.
zinc nitrate. The protein concentration in the absence tion in the case of lead –carbohydrate system,
of zinc remains at 13 ppm in the entire pH range although a distinct adsorption maximum is not
studied. It may be recalled that the protein adsorption observed in this case.
onto sphalerite was found to be higher below pH 6
and very little adsorption was observed beyond pH 10 It is well known that sulphide minerals get hy-
(Fig. 2). Thus, the results of the co-precipitation tests droxylated as a function of pH. In this context, it is
are in agreement with the adsorption data. pertinent to recall that the solubility product of lead
Fig. 10(b) shows the variation in the residual hydroxide is over three orders of magnitude less than
concentration of lead species and protein as a function that of zinc hydroxide (Lide, 1999). Further, zinc
of pH. The lead concentration is marginally reduced hydroxide is amphoteric while lead hydroxide is
in the presence of the metabolite below pH 4, and highly basic (Phillips and Williams, 1965). Thus in
appreciably in the pH range of 10.5 – 11.5, compared the alkaline pH range, zinc hydroxide decomposes to
to that in the absence of the metabolite. The protein ZnO22 ions, while lead hydroxide is stable. The
concentration in the presence of lead nitrate is signif- interaction of polysaccharides with various sulphides
icantly reduced in the pH range of 2– 9, attesting to and oxides has been extensively studied by Laskowski
strong interaction. Such a trend corroborates the and co-workers, and a chemical complexation mech-
adsorption results, wherein higher adsorption of the anism has been proposed (Liu and Laskowski,
protein onto galena was observed below pH 7 (Fig. 2). 1989a,b,c). More recently, an acid – base interaction
model has been suggested by them (Liu et al., 2000).
3.8. Adsorption mechanisms of carbohydrate from B. These mechanisms have been corroborated by the
polymyxa metabolite at mineral surfaces work of Rath and Subramanian (1999a,b), Rath et
al. (2000) for the interaction of polysaccharides such
From the results of the adsorption, flotation, floc- as dextrin and guar gum with several sulphides.
culation and co-precipitation experiments carried out In line with the proposed mechanisms, the hydrox-
on sphalerite and galena, using the bacterial carbohy- yl groups in polysaccharides interact with the mineral
drate, the following facts emerge: surface metal hydroxide both by hydrogen bonding
and chemical forces.
1. The adsorption density of carbohydrate onto galena According to the acid – base interaction model the
is higher than that onto sphalerite. Additionally, the hydroxylated mineral surface would behave as a
amount adsorbed onto galena continuously in- Bronsted base with the hydroxyl groups in the poly-
creases with increase of pH. However, in the case saccharides behaving as a Bronsted acid (Liu et al.,
of sphalerite, an adsorption maximum is observed 2000). It can be expected that stronger the basicity, the
around pH 7. greater will be the interaction with the polysacchar-
2. Flotation and flocculation tests reveal depression of ides. It is thus logical to expect that lead hydroxide
galena and dispersion of sphalerite respectively, in being strongly basic, will have a higher affinity for the
the presence of the bacterial metabolite. bacterial carbohydrate, compared to hydroxylated
3. Co-precipitation tests confirm strong interaction sphalerite, which is amphoteric. The basicity of the
between Zn species and carbohydrate at around pH surface hydroxyl groups will be dependent on the
7, while in the case of Pb species significant valence states, ionic radii and coordination numbers of
interaction is observed in the pH range of 7– 11. On the lead and zinc ions on the corresponding mineral
a comparative basis, the interaction between lead surfaces. For equivalent valence states and coordina-
species and the carbohydrate is significantly higher. tion numbers, ionic radii of zinc are less than that of
4. It is noteworthy that the pH of maximum lead (Lide, 1999). Consequently, the surface hydrox-
adsorption of carbohydrate onto sphalerite coin- ide groups on galena exhibit more basicity resulting in
cides with the pH of maximum co-precipitation for stronger interaction with the carbohydrate. This facil-
the corresponding zinc –polymer system. Similarly, itates selective depression and flocculation of galena
a close parallelism exists between the pH of from sphalerite especially in the alkaline pH range. In
maximum adsorption and significant co-precipita- the case of bacterial carbohydrate, apart from the
S. Subramanian et al. / Int. J. Miner. Process. 72 (2003) 175–188 187
hydroxyl groups, the carboxylate ions, which are between the negatively charged protein molecules and
present in uronic acid could partake in the adsorption the anionic mineral surfaces. Thus, electrostatic, hy-
process. It is understandable that the carboxylate ions drogen bonding, hydrophobic and van der Waals
will be relatively much less compared to the hydroxyl interactions contribute to the adsorption mechanisms,
groups. The presence of carboxyl groups in the in addition to factors leading to conformational
metabolite has been confirmed by FTIR spectroscopic changes of the protein molecules.
studies detailed elsewhere (Santhiya, 2001).
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