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Adsorption of heavy metals

by green algae and ground
rice hulls
a a
Dipak Roy , Paul N. Greenlaw & Barbara S.
b
Shane
a
Department of Civil Engineering, Louisiana
State University, CEBA Building, Baton Rouge,
Louisiana, 70803
b
Institute of Environmental Studies, Louisiana
State University, Atkinson Hall, Baton Rouge,
Louisiana, 70803
Version of record first published: 15 Dec 2008.

To cite this article: Dipak Roy , Paul N. Greenlaw & Barbara S. Shane (1993):
Adsorption of heavy metals by green algae and ground rice hulls, Journal
of Environmental Science and Health . Part A: Environmental Science and
Engineering and Toxicology: Toxic/Hazardous Substances and Environmental
Engineering, 28:1, 37-50

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 J. ENVIRON. SCI. HEALTH, A28(l), 37-50 (1993)

ADSORPTION OF HEAVY METALS BY GREEN ALGAE

AND GROUND RICE HULLS

Downloaded by [State University NY Binghamton] at 23:42 24 April 2013

Key words: Adsorption, heavy metals, green algae, ground rice hulls

Dipak Roy¤, Paul N. Greenlaw¤, Barbara S. Shane*

¤
Department of Civil Engineering
CEBA Building
Louisiana State University
Baton Rouge, Louisiana. 70803
* Institute of Environmental Studies
Atkinson Hall
Louisiana State University
Baton Rouge, Louisiana. 70803

ABSTRACT

This research demonstrates the applicability of low cost, readily available alternate

adsorbents to remove and recover toxic heavy metals from water. Heavy metal ion

adsorption has been investigated using two different adsorbing biomasses, algae and rice

hulls. Algal biomass adsorption studies were conducted with : As, Cd, Co, Cr, Pb, Ni and

Zn, and rice hull biomass adsorption studies were conducted with: As, Cd, Cr, Pb and Sr.

Both biomasses were dried and pulverized to maximize surface area. The green alga,

Chlorella minutissima, adsorbed greater than 90% of the initial Pb, and greater than 98% of

the initial Co concentrations. Rice hulls (Mars and Rico Varieties) exhibited similar

adsorption percentages: 94% for Sr, and greater than 99% for Pb. The rate of adsorption

was fast; the solution metal concentration reached equilibrium within minutes. The cell wall

metal complex was found to be stable; the bound metal did not desorb over time under static

37

Copyright © 1992 by Marcel Dekker, Inc.

 38 ROY, GREENLAW, AND SHANE

conditions. Most cationic metal ions could be recovered from the biomass through

desorption by lowering the pH of the medium.

INTRODUCTION

The problem of heavy metal contamination in waterbodies is widespread and becoming

more acute, threatening an ever increasing portion of the global population. The presence of
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toxic heavy metals in water resources poses unacceptable chronic and acute health risks.

Effective, low cost methods of removing the heavy metal burden of water resources are

urgently needed to restore potability and purity to many water resources.

Chelation of metal ions onto biomass such as cell walls and subsequent recovery, through

the washing of the biomass-metal complex, has been proposed by many researchers as an

effective method of combating the problem of heavy metal contamination. The chelation

technique could be used to reduce the metal concentrations of many industrial effluents, or

for recovery of precious metals in the mining industry. The use of microorganisms,

especially algae, is an obvious choice because of their large surface area and their chelating

potential. Algae are native to a vast array of freshwater and marine environments, and can be

grown in large quantities with relative ease. More importantly, the technique of metal

removal by chelation to cell walls of the biomass has the ability to reduce metal

concentrations in aqueous solutions to 1 ppm or less.

The use of microorganisms in vivo to immobilize metal ions has been used successfully

by many investigators1'2'3. Many of these experiments have demonstrated the potential of

living cells to sequester metal ions from aqueous media. The cell walls of the organisms have

been shown to chelate the majority of the metal ions, either on their cell wall surfaces or

within their cell wall matrices.

Many different types of biomass have been shown to adsorb heavy metals4'5'6'7'8'''10-".

Research on the formation of metal-ligand complexes has suggested that the mechanism of

binding of heavy metals to cell walls of biomass is likely to be independent of metabolism.

 ADSORPTION OF HEAVY METALS 39

Therefore, it is not necessary to use live cells for removal of heavy metals. It is believed

that exposed functional groups within the skeletal structure and surface of the cell wall

coordinate and complex the metal ions. The mechanism of this interaction is unclear,

however. In this research, a green alga and another form of biomass, rice hulls, were

compared for their ability to chelate and remove heavy metals from the aqueous phase. Rice
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hulls and green alga were used because their cellular structure is similar (in terms of

functional group availability), and because of their worldwide availability and abundance.

MATERIALS AND METHODS

Algae: The green alga Chlorella minutissima was obtained from the Phycological Collection

at the University of Texas at Austin, and maintained in a commercial sea salt water medium

("Sea Salt", Lake Products, Ballwin, MO.). The algae were prepared by washing the cells

three times with deionized water, followed by centrifugation at 4,000 rpm for 10 minutes to

concentrate the cells into a soft pellet. The pellet was dried overnight at 60°C.

Rice Hulls: Rice hulls (Mars and Rico Varieties) were obtained from the Louisiana State

University Experiments Station in Crowley, Louisiana. The rice hulls that were used for the

experiments were leftover, unusable by-product from the commercial harvesting of the

Experimental Station's rice crop. The dry rice hulls were ground with a Wiley mill.

Heavy Metal Stock Solutions; 1,000 mg/L - 10,000 mg/L stock solutions of individual

heavy metals were prepared using reagent grade metal chloride or nitrate salts and deionized

water. The pH of the each solution was adjusted to 7 with either HCL or

tetramethylammonium hydroxide.

PIPES Buffer: A 5mM solution of piperazine N N' bis (2 ethane sulphonic acid) buffer (pH

7) was used to resuspend the biomass. Previous research has suggested the use of this buffer

or tetramethylammonium hydroxide for the upward adjustment of pH because neither of these

compounds chelate metals'2.

 40 ROY, GREENLAW, AND SHANE

Analysis of Heavy Metals; The heavy metal analyses were performed in accordance with the

the Method No. 200.7 approved by EPA13. The instrument used was an inductively coupled

argon plasma (ICAP) Jarrell-Ash Atom Comp Series 855 spectrophotometer.

Batch Adsorption/Pesorption Experiments: Batch adsorption experiments were conducted

in five reactors containing only one heavy metal in each reactor. The initial concentration of
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the heavy metals, Cd, Zn, Pb, Ni, and Co were 0.5948, 2.886, 34.48, 70.74, and 164.6

mg/L, respectively, for the individual metal adsorption tests using algal biomass. For

experiments to study adsorption of heavy metals in mixture, individual cationic metal

concentrations were approximately 1 and 40 mg/1, respectively, for algal and rice hull

biomass tests. Anionic species Cr2O7"2 and H2As04' in the metal mixture were present at

approximately 840 and 100 mg/1, respectively, for algal biomass experiments. Approximately

350.0 mg of dry weight of algal biomass was added to 100 ml PIPES (pH 7), in the Teactors.

A pH 7.0 was selected, because for many metals the fraction bound by biomass is maximum

at pH 6 and then remains constant at increasing pH.14 An algal concentration of 3.5 mg/mL

was chosen because it falls within the optimal range of 0.4 mg/mL to 10 mg/mL cited in the

literature151'6. Different studies have shown that increasing the algal cell mass concentration

to 4.5 mg/mL or above did not result an increase in the fraction of metal bound by the

biomass.17 The metal-biomass solution was agitated on a shaker table for the duration of the

test, except during brief periods of sampling. The batch adsorption tests were conducted at

room temperature for at least 30 minutes, which is three times longer than the equilibrium

time of approximately 10 minutes reported in previous experiments.7 Samples were

withdrawn from the supernatant phase, which established itself quickly upon quiescence.

These samples were acidified to pH 2 and stored for analysis.

RESULTS AND DISCUSSION

A similiar series of experiments were performed using algae and rice hulls except the

adsorption/desorption of Co and Zn were undertaken with the algae and the adsorption of Sr

was studied with rice hulls.

 ADSORPTION OF HEAVY METALS 41

The results of the single metal batch adsorption experiments by algal biomass are shown

in Figure 1. The metal concentrations in each solution is normalized to its respective initial

concentration. The rate of adsorption of Cd, Pb and Zn by the algal biomass was rapid,

reaching equilibrium in less than 3 minutes. The adsorption rate for Co and Ni follows the

same trend, although equilibrium was only reached between 15 to 20 minutes. The percent
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removal (percent reduction of initial metal concentration) of Ni was the lowest at 77%, while

Pb was the highest at greater than 99%. On a molar basis, removal of Cd, Zn, Pb, Ni and

Co was 1.5x10*. 1.2xia5, 4.7x10', 2.65xlOJ, and 7.1x10"" mmoles/mg cell mass,

respectively. It is apparent from these results that the removal on a molar basis increases

with an increase in the initial metal concentration. However, the percent removal of the

individual heavy metals does not follow the same order as the loading rate. To illustrate this

point, consider the findings with Pb and Ni. The initial concentration of Ni (70.74 mg/L) is

almost twice that of Pb (34.48 mg/L), but th& % removal of Ni is 77% while that of Pb is

99%. These results suggest that the site of Ni adsorption is probably different from that of

Pb, and furthermore the number of adsorption sites for individual heavy metals per unit of

biomass is different.

The process of adsorption of heavy metals to cell wall surfaces or within skeletal cell

wall structures may be conceptualized as composed of two steps: (a) transport of heavy metal

ions from the bulk solution through a liquid film to the cell wall surface and to interskeletal

space; and (b) binding of metal ions to specific target sites. A higher initial metal

concentration will result in a higher driving force for transport of cations from the bulk

liquid to the solid surface resulting in a higher adsorption capacity. The adsorption capacity

will level off when all the specific binding sites of the biomass are occupied. In contrast,

many researchers have reported a decrease in binding capacity with increasing adsorbent

concentrations, for the same adsórbate concentrations." To investigate this finding further,

experiments were undertaken in which Cd was added at two different initial concentrations,
 42 ROY, GREENLAW, AND SHANE

TABLE I
Adsorption of two concentrations of Cd by
Increasing Algal Biomass Concentration

Biomass Cd 58.84 mg/L Cd 99.76 mg/L «is/qjs

Cone, (mg/ml) q (mmol/mg) q (mmol/mg)

0.25 0.001952 0.003279 1.67

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0.5 0.000981 0.001641 1.67

1.0 0.000498 0.00083 1.69
3.0 0.00017 0.000284 1.67

58.84 and 99.76 mg/L, with cell biomass concentrations of 0.25, 0.5, 1.0 and 3 mg/mL.

The results of this experiment, summarized in Table 1, show the adsorption capacity, (q), in

mmoles/mg of cell mass at the two initial metal concentrations for the four different algal

loadings. It is apparent from this table that q^qs, is approximately the same at 1.67 for all

combinations of biomass concentrations at the two concentrations of the metal, i.e, this ratio

is same as that of the initial metal concentrations of 99.78 and 58.84 mg/L. For a given cell

mass, an increase in metal concentration resulted in a greater binding of the metal/mg of cell

mass. Over an increasing range of biomass concentration, the adsorption capacity decreases

in the same relative proportion as the biomass concentration increases. Based on the results

reported here, it is difficult to predict the saturation level of q, which is a function of the

type of biomass, the metal ion species, pH and other operating conditions. Thus additional

experiments must be conducted to establish the saturation level before this process can be

adopted for "real world" applications.

To ascertain whether the adsorption of heavy metals by algal biomass was competitive

or noncompetitive, additional batch adsorption experiments were undertaken using a mixture

of Cd, Ni, Pb and Zn salts at individual metal concentrations of 1 mg/mL. Figure 2 shows a
 ADSORPTION OF HEAVY METALS 43
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i 10 20 30 40 50 60 70

Time (minutes)

Figure 1: Adsorption of Cd, Co, Ni, Pb and Zn by algal biomass from single metal solution.

10 20 30 40 50 60 70 80

Tims (minutes)

Figure 2: Adsorption of Cd, Ni, Pb and Zn by algal biomass from metal mixture solution.
 44 ROY, GREENLAW, AND SHANE

TABLE 2
Comparison of the Adsorption Capacity, (inmole Metal/mg Biomass) of Algae
for Four Metals added in mixture.

Metal Ratio of Observed Predicted Observed

initial [Me] adsorption adsorption adsorption
Mixture/Single capacity in capacity for capacity for
single test mixture test mixture test
q, q„p
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Cd 1.632 1.5x10* 2.45x10* 2.4x10*

2 4
Ni 1.387X10- 2.65X10- 3.68x10* 4.7x10*
2 5
Pb 2.859xlO 4.7xlO 1.34x10* 1.3x10*
Zn 3.274x10' 1.2xlO5 3.93x10* 3.7x10*

rapid binding of all four metals with similar trend to that shown in Figure 1. If one compares

the results from this experiment to that in which adsorption of the single metals were studied,

it is possible to determine the adsorption capacity of each specific metal by the algal

biomass. The adsorption capacity values from the tests with single metal were used to

predict adsorption capacity of the algae for the metals when tested in the mixture. This metal

mixture adsorption value, (qaj, was obtained by multiplying the adsorption capacity of the

algae for a metal, ( q j , by the ratio of the initial metal concentration in the two tests ([Metal]

concentration for the mixture test/[Metal] concentration for the individual metal test). The

values, ta.»), are the adsorption capacity of the algae for the individual metals when tested as

a mixture with the algal biomass. The adsorption capacities of the algal biomass are shown in

Table 2. The close agreement between the predicted and observed adsorption capacities

suggests that the adsorption process is noncompetitive. There is less than a 1% difference

between the predicted and observed adsorption capacity values for Cd, Pb, and Zn but with

Ni the predicted value is nearly 22% less than that observed. The latter observation may be

due to experimental errors.

 ADSORPTION OF HEAVY METALS 45

50
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10 20 30 40 60 60 70

Time (minutes)

Figure 3: Adsorption of Cd, Pb and Sr by rice hull biomass from metal mixture solution.

The next series of experiments were performed with ground rice hulls as the biomass

adsorbent. The results of the adsorption experiment with a solution of Cd, Pb and Sr are

shown in Figure 3. As with the adsorption experiemnt with the algal biomass, a rapid

equilibrium was established with the rice hull biomass. The respective adsorption of Pb, Cd,

and Sr was 99.43%, 97.96%, 94.11% of the initial metal concentration.

Thus far, the results presented describe the adsorption of divalent cations only.

Previous investigations have also concentrated on studies of divalent cation adsorption.

Heavy metals such as As and Cr, which are present in the natural environment in anionic

forms are also of concern due to their high toxicity. Figure 4 shows the results of adsorption

of anionic species Cr2O7"2 and H2As04' on algal biomass. As seen in the case of cation

adsorption, the equilibrium level for anion adsorption on algae was also reached within

minutes. Adsorption of 97.95% of the added chromium and 97.91% of the added arsenic to
 46 ROY, GREENLAW, AND SHANE

110

100
3
90
.As
eo
"5 _Cr
c
o 70

60
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50
8
40
1
i 30

20

10

0
10 20 30 40 50 60 70 60

Time (minutes)

Figure 4: Adsorption of anionic As and Cr by algal biomass from metal mixture solution.

the biomass was observed. Similar experiments with rice hull biomass and anionic metal

species show 98.93% and 98.95% adsorption of Cr and As, respectively (data not shown).

Table 3 shows a comparison of the adsorption capacities of both algae and rice hulls, of

two cations Pb and Cd, and two anions Cr2Cy2 and HjAsO4'. Since the initial metal

concentrations used in the two cation mixture tests (algae and rice hulls) were different, a

ratio of the initial, individual metal concentrations (initial [Metal] in the metal mixture test

with the rice hulls/ initial [Metal] in the metal mixture test with the algae) was used to

predict the adsorption capacity of the algal biomass at the initial metal concentration that was

tested with the rice hulls. It is apparent from the comparison presented in Table 3 that rice

hull biomass is as good an absorbent as algae for removal of As, Cr and Pb. However, for

Cd, adsorption capacity of rice hulls is twice as high as that of algae.

 ADSORPTION OF HEAVY METALS 47

TABLE 3

Comparison of Adsorption Capacity (mraoles/mg)

of the Algae and Rice Hulls

Metal Initial Metal Predicted Algal Observed rice

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Cone, in Rice adsorption adsorption

Hull Expt., mg/1 capacity capacity

Pb 40.0 5.30xl0 5 5.5xl0"5

Cd 40.0 9.91xlO s 1.9X10"4

CrA" 3 410.0 3.12xlO J 3.16xl0 J

H 2 AsO 4 ' 241.0 8.12x10» 8.21xl0 3

Application of biomass to remove heavy metals from aqueous solutions could be a useful

process provided there is an efficient and economical method of regenerating the spent

biomass. One of the most common methods of desorption of heavy metals from biomass is

by adjusting the pH. In these experiments, the desorption was initiated by reducing the pH

of the biomass samples loaded with heavy metals to 1.55 with concentrated HCL. Figure 5

shows the results of two independent desorption experiments conducted on the algal and rice

hull biomass. The bar graph illustrates the increase in metal concentration before and five

minutes after the solution pH was lowered to 1.55. The percent of adsorbed metal that

desorbed from each biomass is quite variable for each metal. Metal desorption from the

algal biomass was high for Pb and Zn at 90.2% and 91.5%, respectively, and low for Ni and

Cd at 1.2% and 0.3%, respectively. Metal desorption from the rice hull biomass was low:
 48 ROY, GREENLAW, AND SHANE

6 1
5.5 (A) Ground rice hulls
I 5 -
4.5

i
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4
3.5-
!
3 •
8
2.5
2
1.5
1 .
0.5
0
Lead Cadmium Strontium

0.6
0.55 Legend
0.5 (B) Green algae
i Equil. soin (M) cone
Í3S 0.45 Desorb soin (M) cone
~ 0.4
f 0-35

8 °"3
8 °25
§ 0.2
| 0.15
| 0.1
< 0.05

Lead Cadmium

Figure 5: Desorption of cationic heavy metals from rice hull and algal biomass.
 ADSORPTION OF HEAVY METALS 49

9.5%, 6.9%, and 5.2% for Cd, ST, and Pb, respectively. The desorption of the metals

from the biomass upon a pH drop is in accordance with reported results on desorption of

metal bound to algae1-14. In this research, full regeneration was not attempted; however, the

volumes of concentrated HCL used were quite small, less than 3 mL for 350-500 mg of

biomass and desorption time allowed was 5 minutes only.

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CONCLUSIONS

This study has demonstrated that dried Chlorella minuiissima cells and ground rice hulls

are good heavy metal adsorbents. The specific findings are summarized below:

1. Metal adsorption by algal and rice hull biomass, from the aqueous test systems, was

greater than 90% for all the metals tested, (Sr, Cd, Ni, Pb, Zn, Co, Cr, As), except Ni, for

which removal was nearly 80%.

2. Adsorption equilibrium was established rapidly, occuring within minutes.

3. Adsorption of heavy metals by algal biomass was observed to be noncompetitive.

4. Adsorption capacity for ground rice hulls was comparable to that observed for the algal

biomass.

5. Both the ground rice hulls and the algal biomass were found to desorb cationic metal ions

upon lowering the solution pH to 1.55.

REFERENCES

1. Fisher, N.S. Bioaccumulation of technetium by marine phytoplanton. Environ. Sci.

Tech. 1982; 16: 579-581.

2. Norberg, A.B. Accumulation of heavy-metal ions by Zoogloea ramigera. Biotechnol.

Bioeng. 1984; 26: 239-246.

3. Wang, H.K. Bioaccumulation of nickel by algae. Environ. Sci. Tech. 1984; 18: 106-
109.

4. Tsezos, M., Volesky, B. The mechanism of uranium bioadsorption. Biotechnol.

Bioeng. 1982; 24: 385-401.

5. Watkins, J.W., Elder, R.C., Greene, B., Darnall, D.W. Determination of gold binding in
an algal biomass using EXAFS and XANES spectroscopies. Inorg. Chem. 1987; 26:
1147- 1151.
 50 ROY, GREENLAW, AND SHANE

6. Jennett, J.C., Hassett, J.M., Smith, J.E. Control of heavy metals in the environment
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8. Gale, N. and Wixson, B. Removal of heavy metals from industrial effluents by algae.
Develop. Industr. Microbiol. 1979; 20: 259-273.
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Hydrogen ion buffers for biological research. Biochem 1966; 5(2): 467-477.

13. EPA. Inductively Coupled Plasma-Atomic Emission Spectrophotometric Method for

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14. Darnall, D.W., Greene, B., Henzl, M.T., Hosea, M., McPherson, R.A., Sneddon, J.,
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Environ. Sci. Tech. 1986; 20: 206-208.
15. Standberg, G.W., Shumate II, S.E., Parrott Jr., J.R. Microbial cells as biosorbents for
heavy metals: Accumulation of uranium by Saccharomyces cerevisiae and Pseudomonas
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16. Greene, B., McPherson.R., Darnall, D.W. Algal sorbents for selective ion recovery. In:
Metals Speciation, Separation, and Recovery. 1987; Lewis Publishers Inc.

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Date Received: 06/09/92

Date Accpeted: 07/13/92