UNIVERSITI PUTRA MALAYSIA REMOVAL OF Cu (II) AND Cr(VI) FROM AQUEOUS SOLUTIONS BY WATER LILIES (NYMPHAEA SPONTANEA) CHOO TZE PEI FSAS 2001 27
UNIVERSITI PUTRA MALAYSIA
REMOVAL OF Cu (II) AND Cr(VI) FROM AQUEOUS SOLUTIONS BY WATER LILIES (NYMPHAEA SPONTANEA)
CHOO TZE PEI
FSAS 2001 27
REMOVAL OF Cu (II) AND Cr(VI) FROM AQUEOUS SOLUTIONS BY
WATER LILIES (NYMPHAEA SPONTANEA)
By
CHOO TZE PEI
Thesis Subm itted in Fulfilment of the Requ irement for the Degree of Master of Science in Institute of Bioscience
Universiti Putra Malaysia
June 2001
iii
Abstract of thesis presented to the Senate ofUniversiti Putra Malaysia in fulfilment of the requirements for the degree of Master of Science.
REMOVAL OF Cu(Il) AND Cr(VI) FROM AQUEOUS SOLUTIONS BY WATER LILIES (NYMPHAEA SPONTANEA)
By
CHOOTZEPEI
June 2001
Chairman: Professor Lee Chnoong Kheng, Ph.D
Faculty: Science and Environmental Studies
This study describes an investigation using water lilies (Nymphaea spontanea) to
remediate heavy metals (Le. chromium and copper) from aqueous solutions and
electroplating waste. Laboratory tests were conducted to evaluate the effect of
different ages of water lilies in removing chromium and the efficiency of water lilies
in accumulating different metal ions (i.e. chromium and copper) at various
concentrations. Phytotoxicity tests on the test plants were also conducted to
determine the plant tolerance towards these metal ions.
The absorption of metal ions by aquatic plants depends on the nature and the
amount of aquatic plants, their stage of development, as well as the metal ion
content and the presence of other dissolved substances. This study shows that plant
age has significant effect on the removal and accumulation of chromium. Plants of 9
IV
weeks old appeared to accumulate the most chromium followed by 6 and 3 weeks
old plant. The roots of water lilies play an important role in the uptake of metal ions
and the results show that metal accumulation in water lilies follow the order: roots >
leaves> petioles. Water lilies are capable of accumulating substantial amount of
chromium. up to 5956 �g, and copper, up to 10615 �g, and it is shown that
copper is removed more rapidly than chromium. The preferential accumulation of
copper over chromium is probably due to the selective nature of these plants and the
chemical behaviour of the metal ions. Removal of metals by water lilies is more
efficient when the metal is present singly in the solution than in mixed metal
solution or waste solution. Results also show that copper is more toxic to water lilies
when compared to chromium.
Abstrak tesis yang dikemukakan kepada Senat Universiti Putra Malaysia sebagai memenuhi keperluan untuk ijazah Master Sains.
PENYINGKIRAN Cu(ll) DAN Cr(VI) DARIP ADA PELBAGAI LARUT AN AKUAS OLEH BUNGA TERATA! (NYMPHAEA SPONTANEA)
Oleh
CHOO TZE PEl
Jun 2001
Pengerusi ; Profesor Lee Chnoong Kheng, Ph.D
Fakulti: Sains dan Pengajian Alam Sekitar
v
Penyelidikan ini bertujuan untuk mengkaji potensi bunga teratai (Nymphaea
spontanea) dalam bioremidiasi buangan logam berat umpamanya kromium dan
kUprum. Ujian makmal telah dijalankan bagi menilai kesan peringkat kematangan
bunga teratai dalam remidiasi kromium dan kecekapan bunga teratai untuk
menyerap dan mengumpul pelbagai jenis ion logam dalam kepekatan yang
berlainan. Ujian tosikologi logam ion terhadap bunga teratai juga dikaji bagi
menentu ketahanan I toleransi tumbuhan terhadap logam-Iogam ion ini.
Penyerapan logam ion oleh tumbuhan akuatik adalah bergantung pada faktor-faktor
seperti peringkat kematangan atau pertumbuhan dan logam ion yang didedahkan
pada tumbuhan tersebut dan juga bahan yang terlarut dalam efluen buangan logam.
Kajian ini telah menunjukkan peringkat kematangan tumbuhan yang berlainan
vi
mempunyai keupayaan penyerapan dan pengwnpulan logam ion yang berlainan.
Tumbuhan yang berusia 9 minggu didapati berupaya mengumpul logam ion yang
banyak sekali, dituruti tumbuhan yang berusia 6 dan 3 minggu. Akar bung a teratai
memainkan peranan yang penting dalam peyerapan logam ion dan didapati dalam
kajian ini, tren pengumpulan logam ion dalam bahagian-bahagian tumbuhan adalah
seperti berikut: akar > daun > batang. Bunga teratai juga didapati berupaya
mengwnpul sebanyak 5965 �glg logam kromium dan 106 1 5 J-Lg/g log am kuprum.
Di samping itu, kajian ini juga menunjukkan bahawa lebih banyak logam kuprum
dapat diremediasi daripada logam kromium. Ini mungkin disebabkan selektiviti
tumbuhan tersebut terhadap logam ion yang tertentu dan juga suat kimia logam ion
tersebut. Kecekapan bunga teratai dalam penyerapan logam ion didapati lebih
rendah dalam larutan yang rnernpunyai pelbagai jenis logam berbanding larutan
yang hanya rnernpuyai sejenis logarn dalarnnya. Keputsuan-keputusan kajian ini
juga mempamerkan bahawa logam kuprum adalah Iebih toksik daripada kromium
terhadap tumbuhan.
Vll
ACKNOWLEDGEMENTS
First of al� I would like to extend my most sincerer gratitude to my project
supervisor, Professor Dr. Lee Chnoong Kheng, co-supervisors, Associate Professor
Dr. Low Kun She and Dr. Hishamuddin Dmar for their guidance, ideas, suggestions
and constructive criticism throughout my research. The sharing of ideas has not only
been helpful in the progress of my project but also served as useful principles of life
in years to come, and these experiences are indeed invaluable.
I would also like to extend my gratitude to all the technical staff of the Chemistry
department especially Madam Choo, and Dr. Mohd. Said Saad, En. Tajuddin of
Plant Genetic Resource Department of Institute of Bioscience for their assistance
and helpful suggestion towards the success of this study. Besides that, I would also
like to thank Dr. Tan Hee Siong for his suggestion on the methodology of some
biochemical analysis and for helping me to get the material for sugar analysis.
I would also like to extend my gratitude to Mr. Ng Kok Hong of Lakescape
Integration for his kindness to teach me the planting and growing skill of water
lilies.
VUl
In addition, I would like to thank my good friends Su Sim and Zaid for landing their
hand in planting water lilies for my research and lab-mates who are also working
under the supervision of Prof. Lee for their support, suggestions and comments
throughout my project. Last but not the least, I would like to thank my family
especially my parents and my brothers Jimmy and Johnny who are my source of
inspiration and strength. Also not forgotten are my friends, especially Chooi Phyng,
Nyuk Y ong, Mei Chee, Tien Siong and my house-mates for their continuous support
and encouragement; without them, I would not have come this far.
1X
I certifY that an Examination Committee met on 7th June 2001 to conduct the fmal examination of Choo Tze Pei on her Master of Science thesis entitled "Removal of Cu(II) and Cr(VI) from Aqueous Solutions By Water Lilies (Nymphcrea spontanea)" in accordance with Universiti Pertanian Malay sia (Higher Degree) Act 1980 and Universiti Pertanian Malaysia (Higher Degree) Regulation s 1981. The Committee recommends that the candidate be awarded the relevant degree. Members of the Examination Committee are a s follows:
Md Jelas Bin Haron, Ph.D. Associate Professor Faculty of Science and Environmental Studies Universiti Putra Malaysia (Chairman)
Lee Chnoong Kheng, Ph.D. Professor Faculty of Science and Environmental Studies Universiti Putra Malaysia (Member)
Low Kun She, Ph.D. Associate Professor Faculty of Science and Environmental Studies Universiti Putra Malaysia (Member)
Hishamuddin Bin Omar, Ph.D. Faculty of Science and Environmental Studies Universiti Putra Malaysia (Member)
MO . HAZAI1MOHAYIDIN, Ph.D. Profess eputy Dean of Graduate School Universiti Putra Malaysia
Date: '1 � ,: , < 20m
x
This thesis submitted to the Senate of Uruversiti Putra Malaysia has been accepted as fulftlment of the requirement for the degree of Master of Science.
AINI IDERIS, Ph.D. Professor lDean of Graduate School Universiti Putra Malaysia
Date:
xi
DEC LARA TION
I hereby declare that the thesis is based on my original work except for quotations and citations which have been duly acknowledged. I also declare that it has not been previously or concurrently submitted for any other degree at UPM or other institutions.
CHOOTZEPEI
Date: '3 I 0" { 0 I
TABLE OF CONTENTS
DEDICATION ABSTRACT ABS1RAK ACKNOWLEDGEl\1ENTS APPROVAL DECLARATION UST OF TABLES UST OF FIGURES
CHAPTER
1
2
3
INTRODUCTION 1.1 The hnportance of Water 1.2 Water Pollution 1.3 Technologies for Wastewater Treatment 1.4 Objectives
LITERATURE REVIEW 2.1 Industrial Wastewater Pollution
2.1.1 Hazardous Waste 2.1.2 Heavy Metals 2.1.3 Electroplating Waste 2.1.4 Chromium 2.1.5 Copper
2.2 New Technology for Heavy Metal Removal 2.2.1 Bioremediation 2.2.2 Bioremediation of Heavy Metals
2.3 Bioremediation Using Plants 2.3.1 Aquatic Plants 2.2.3 Aquatic Plants for Remediation of Heavy Metals
2.4 Water Lily for Bioremediation 2.4.1 Biology of Water Lily
MA TERlALS AND METHODOLOOY 3.1 Preparation of Plant Samples 3.2 Metal Uptake Studies 3.3 Study of the Effect of Plant Maturity on Meta] Uptake 3.4- Bioremediation of Electroplating Waste
xu
Page
11
111
V
vu
IX
xi xiv xv
1.1 1.1 1.2
1.3
2.1
2.1
2.2
2.7 2.8 2.13 2.17
2.17 2.20 2.21 2.23 2.24 2.26
2.27
3.1 3.2
3.4
3.4
3.5 Plant Analysis 3 .6 Chlorophyll Analysis 3.7 Protein Analysis
3.7.1 Reagents 3.7.2 Procedure
3.8 Sugar Analysis 3.8.1 Reagents 3.8.2 Procedure
Xlll
3.5 3.6 3.8 3.8 3.9 3.9 3.10 3.lO
4 RESULTS AND DISCUSSION
5
4.1 Results 4.1 4.1.1 Effectiveness of Metal Removal by Water Lilies of
Different Plant Maturities 4.1
4.1.2 Efficiency of Water Lilies in Removing Different Metal 4.3 Ions
4.l.3 Toxicity Effects of Metal Towards Water Lilies 4.16 4.1.4 Studies of the Uptake and Toxic Effects of Mixed Metals 4.19
of Cr and Cu on Water Lilies 4.1.5 Bioremediation of Electroplating Waste by Water Lilies 4.23 4.1.6 Comparative Study of Metal Removal in Different Systems 4.25
4.2 Discussion 4.28 4.2.1 The Importance Plant Maturity in Metal Absorption 4.28 4.2.2 Mechanism of metal Uptake in Relation to Deposition of 4.29
Metal in Plant 4.2.3 Metal Accwnulation 4.34 4.2.4 Phytotoxicity Effects in Relation to Metal Accumulation 4.38 4.2.5 Bioremediation of Electroplating Waste 4.41 4.2.6 Potential of Water Lilies (N. spontanea) for 4.41
Bioremediation
CONCLUSION 5.1
REFERENCE SJB ffiLIOGRAPHY APPENDICES
R.l A.1 B.l BIODATA OF TIlE AUTIIOR
XIV
LIST OF TABLES
Table Page
2.1 Maximum pennissible concentrations of various metals in Datural 2.3 waters for protection of human health
2.2 Composition of raw waste streams from common metal plating 2.8
2.3 Worldwide anthropogenic input of Cr into freshwater 2.11
2.4 Worldwide anthropogenic input ofCu into freshwater 2.15
4.1 Concentration factors (CF) for Cr and percentage of Cr removed by 4.1 water lilies of different ages after 7 days of exposure
4.2 Effectiveness of water lilies in removing Cr and Cu after 7 days of 4.7 exposure
4.3 Effect of Cr and Cu on selected biochemical parameters of water 4.17 lilies
4.4 Toxicity effect ofCr and Cu on selected biochemical parame ters 4.18 in water lilies 7 days after treatment
4.5 Concentration factors for Cr and Cu and percentage of metal 4.22 removed by water lilies in mixed solutions of different ratios of metals
4.6 Toxicity effect of mixed metal solution on changes in chlorophyll, 4.23 protein and sugar in water lilies on 7th day after treatment
4.7 Toxicity effect of electroplating: waste on selected biochemical 4.26 parameters in water lilies on 7 day after treatment
A.l Biomass of water lilies of different ages A.l
xv
LIST OF FIGURES
Figure Page
2.1 Plant structure of water lily (Nymphaea spontanea) 2.28
4.1 Chromium removal by water lilies of different ages 4.2
4.2 Means of Cr uptake (mg/g) in relation to different ages of water lilies 4.4
4.3 Chromium remaining in solution after treatment by water lilies 4.5
4.4 Percentages of Cr uptake in relation to different metal concentrations 4.6 after treatment with water lilies
4.5 Copper remaining in solution after treatment with water lilies 4.8
4.6 Percentages of Cu uptake in relation to different metal concentrations 4.9 after treatment with water lilies
4.7 Effects of Cr and Cu concentration on metal accumulation by water lilies 4 . 1 0
4.8 Percentages of Cr and Cu uptake in relation to different metal 4.11 concentrations after treatment with water lilies
4.9 The bioaccumulation pattern of chromium ions in different plant parts 4.13 of water lilies
4.10 Accumulation of eu in different plant parts of water lilies 4.14
4.11 Metal concentration in different plant parts of water lilies 4.15
4.12 Percentage of Cr removed in different ratios of mixed solution after 4.20 treatment with water lilies
4.13 Percentage of eu removed in different ratios of mixed solution after 4.20 treatment with water lilies
4.14 Metal accumulation by water lilies after treatment in different ratios of 4.21 mixed metal solution
XVl
4.15 Percentage of metal remaining in waste solution after treatment with 4.24 water l ilies
4.16 Metal accum ulation by water lilies after treatment in electroplating waste 4.24
4.17 Removal of chromium from different synthetic and waste solutions by 4.27 water lilies
4.18 Removal of copper from different synthetic and waste solutions by 4.27 water lilies
1.1 The Importance of Water
CHAPTER 1
INTRODUCTION
1.1
The recent water crisis in Malaysia, a country which is well-known for her water
abundance, has led us to ponder on our mistake in thinking that water is an infmite
resource. Water is an essential element, without which life cannot be sustained. In
fact, water is uniquely fit as an environment for living things and provides millions
of organisms with nourishment all the time. Thus, the role of water in nature is
obvious. Man cannot live without water. Although there is growing recognition that
instituting better water control is needed and that global water resources are limited
and are rapidly becoming polluted, the full awareness of water as a precious and
active element in nature has yet to be awakened.
1.2 Water Pollution
Water pollution has significantly become a major global problem. Concerns arise
when man starts to realize that their "used to be" clean water is no longer safe to
drink or play with. Contamination of water usually originates from pesticides runoff
from agricultural land, industrial discharge into sutface waters and urban runoff.
These pollutants when interact with water may produce harmful effects to human,
animal and plants.
1.2
The .Minamata disease caused by mercury contamination in fIsh and the itai-itai
disease due to ingestion of cadmium in water and food that occurred in Jap� are
examples of severe health hazards caused by water pollutants (Yao, 1984). In
addition, some of these pollutants are mutagenic and / or carcinogenic or have
serious ecological implications (Esch, 1977).
1.3 Technologies for Wastewater Treatment
Generally water contaminants are subjected to treatments which involve reduction
or elimination of suspended solids and particulate matter, both organic and
inorganic . These processes also remove substances which are of nutritive value,
such as nitrogen and phosphorous, as well as those that are toxic to organisms, such
as heavy metals, pesticides and other organic compounds.
Presently, the main technologies which have been utilised to reduce or eliminate
these pollutants include lime precipitation, ion-exchange, adsorption onto activated
carbon, membrane processing and electrolytic methods. However , these
conventional physical and chemical treatment procedures have been found to be
limited, since they often involve high capital and operational costs. Furthennore,
these methods are normally associated with the generation of secondary wastes
which present treatment problems, for example , the large quantity of sludge
generated by precipitation process (William et al., 1998).
1.3
Nevertheless, the increasing demand for an improved environment has resulted in
the search for better and cheaper wastewater treatment system. Bioremediation, a
process that employs biological activity to mineralize organic contaminants or
remove inorganic contaminants from polluted sites, has been gaining wide spread
attention.
In this study, water lily (Nympheae spontanea), a natural filter in the aquatic system
was tested for its potential for removing pollutants in industrial wastewater.
1.4 Objedives
The objectives of this study were as below:
To determine the effectiveness of heavy metal removal by water lily.
To study the effect of heavy metals towards plant growth.
To study the effect of plant maturity on its removal of heavy metals.
To evaluate the effectiveness of using water lily in the treatment of industrial
wastewater.
CHAPTER 2
LITERATURE REVIEW
2.1 Industrial Wastewater Pollution
2.1.1 Hazardous Waste
2.1
Wastewater discharged by industrial operations are in some cases among the worst
sources of water pollution. These pollutants are nonnally associated with the
problems of severe oxygen depletion and / or turbidity, in other word, high BOD
(biological oxygen demand), sedimentation or high concentration of suspended
solids and the presence of toxic substances or hazardous waste (Laws, 1993).
Hazardous waste production is a by-product of industrialization. With the increase
in emphasis on high technology industries, hazardous waste production has become
more significant and the range of wastes produced becomes more varied. In
Malaysia, it was found that the total quantity of various types of hazardous waste in
1987 was 380,000 cubic meter per year, and this amount was predicted to be tripled
or more in the coming years ( ShafIi, 1992).
Hazardous wastes which contain heavy metals have aggravated the problem of
water pollution. The problem becomes complex when several metal ions are
involved, fonning metal chelation resulting in unpredictable physical and biological
2.2
changes in the environment according to the industries involve. In fact, the damage
is more direct, since poisons are involved.
2.1.2 Heavy Metals
Heavy metals are defmed as metals which show a specific gravity> 4.5 glroL
(Outridge and Noller, 1991); or elements which under biologically significant
conditions tend to exist as cations (Phipps,1976). However, this term is often used
where there are connotations of toxicity. Cadmium, copper, nicke� lead, chromium
and mercury are most important heavy metal water pollutants. Some of these heavy
metals (e.g. copper, zinc and iron) are essential micronutrients to plants and human,
but others such as mercury, lead and cadmium have no known biological function.
Virtually, all metals including the essential metal micronutrients, are toxic to aquatic
organism as well as humans, if exposure levels are sufficiently high. Table 2.1 gives
some idea of relative toxicity of various metals based on the maximum permissible
concentrations in water recommended by the Environmental Protection Agency
(EPA) for the protection of human health.
Since heavy metals are elements, they cannot be degraded once released. Despite
this, heavy metals are not environmentally "constant" when released, but rather may
be affected in several ways. For instance, biological and physical forces in the
environment can change the chemical fOIm of the heavy metals. This means
2.3
interconversion between organic and inorganic fonns of the metals, or changes in
the organic fonns. Due to these changes, there is a marked difference in the
reactivity of the heavy me tals with biological and non-biological sites in the
environment, thus resulting in uncertainties for scientis ts and regulatory officials in
assessing the environmental consequences of increase in the heavy metal burden.
Table 2.1 Maximum permissible concentrations of various metals in natural
waters for protection of human health
Maximum Permissible Concentration
Metal Chemical symbol mglL J.!MIL
:tviercury Hg 0.144 0.72
Lead Pb 5 24
Cadmium Cd 10 89
Selenium Se 10 127
Thallium Tl 13 64
Nickel Ni 13.4 228
Silver Ag 50 464
Manganese Mn 50 910
Chromium Cr 50 962
Iron Fe 300 5372
Barium Ba 1000 7281
Sow-ce: EPA (1987)
2.4
TIle fact that heavy metals have been present in some parts of the environment for
millennia is recognized. Metals are introduced naturally into aquatic system as a
result of the weathering of soils and rocks, and from volcanic eruptions. However, a
more important source of increased heavy metal concentration is the by-products of
modem society. Mining and mineral processing, fossil fuel combustion, the
dumping and burial of industrial waste, car fumes, domestic waste discharge are few
examples of ways in which modern society may alter the environment (Manahan,
1994).
2.1.2.1. The Toxicity and Effect of Heavy Metals
The toxicity of heavy metals, manifested in aqueous solution may be expressed
through acute poisoning, teratogenicity, mutagenicity, or carcinogenicity (Stoner,
1993). A metal toxicant may bring about one or more of these effects. If more than
one is exhibited, then the effects are likely dose dependent. Metal toxicity may
affect all forms of life including microorganisms, plants and animals, but the degree
of toxicity varies for different organisms.
The effect of heavy metals on the environment depends on the concentrations of the
heavy metals and their chemical forms. Heavy metals are particularly toxic in their
chemically combined forms and some, notably mercury, are toxic in the elemental
form (Manahan, 1994). For a single element, the concentration of direct importance
2.5
may be that :fraction of the total which is available to be mobilized by physical
forces such as wind, water, or soil, and the partition of this fonn among the physical
and biological constituents of the environment
Even when the concentration of metal toxicant may be below the permissible
concentration, the presence of this substance may, nevertheless, constitute a health
hazard due to the phenomenon of bioaccumulation (Stoner, 1993). A metal toxicant
may be accumulated and concentrated within a single organism by ingestion over
time or through successive trophic levels in a food chain. These cwnulative effect
are dependent on the fixation of the toxic substance in the organism involved or on
the absence of significant excretion at any trophic level. In addition, chemical
interconvertion also contributes to significant change in the concentration of the
metal in a given locale. Unavoidable mobilization of heavy metals that results from
leaching of ores by rain or running water is a good example to describe this
occurrence.
Increasing contamination of the aquatic environment by heavy metals posed a
serious threat to biota (Forstbner and Wittmarm, 1979). The presence of heavy
metals in the environment can be detrimental to a variety of living species, including
man. Heavy metals produced undesirable effects, even if they were present in