THE EFFECT OF PH AND AGITATION TIME ON LEAD BIOSORPTION EFFICIENCY FROM AQUEOUS SOLUTION BY DUCKWEED (Lemna sp.

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A Research Paper Presented to the School of Technology University of the Philippines Visayas Miag-ao, Iloilo

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In Partial Fulfilment of the Requirements in ChE 198

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By Juniper V. Magallanes, Jerome T. Magdato and Alyssa Jean C. Sala October 2011

ABSTRACT

Different technologies have been developed with the growing concern for environmental protection. Phytotechnology, the use of plant in combatting environmental issues, have been the focus of some studies for the past years. Duckweeds (Lemna sp.) collected from UPV Facultative Pond were used in the experiment as biosorbent in the removal of lead (Pb) from aqueous solution. The experiment was performed using different pH levels (4.0, 6.0, 7.0, 8.0 and 10.0) with 45-minute and 90-minute agitation times. The result showed the highest percent removal for the pH value of 10.0, 79.7% and 79.9%, at 45 minute and 90 minute agitation time, respectively. The 90- minute agitation time gave higher % removal of lead compared to the 45-minute agitation time at the same pH level. As can be clearly seen in the result, the basicity or acidity of the solution affects the amount of lead removed from it. Generally, the amount of lead removed from the solution increases as the pH level of the solution increases. The result shows a great potential of duckweed as biosorbent material in the removal of heavy metals in wastewater.

Keywords: Duckweed, lead removal, biosorption, pH level, agitation time.

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ACKNOWLEDGEMENT "The acomodador or giving-up point: there is always an event in our lives that is responsible for us failing to progress: a trauma, a particularly bitter defeat, a disappointment in love, even a victory that we did not quite understand, can make cowards of us and prevent us from moving on." Paulo Coelho, The Zahir In accomplishing our study, we had not only been through one but many giving up points. Sometimes, it made us stop and think twice about whether we had made the right choices. Other times, it had almost led us to stray from the right track we're supposed to take. However, looking back, we realized that unlike most people, we have reached, got over through our acomodadors, and turned each into another spark of hope. With strong will, eager minds and unconditional support from every good soul we know, we have survived each point. To our parents, for their undying support and endless love even when our biddings often leave their pockets empty, thank you and we surely love you more. To our classmates, for brightening up our grumpy moods by cracking the corniest jokes on earth, for almost five years of happy days and chismisan sessions, and for letting us know we're never alone, our deepest gratitude and big bear hugs, at last friends, we did it! To all our friends, for putting up with our stress-provoked-monster attitudes, for never hesitating to lend their shoulders and ears whenever we have troubles and most importantly for sharing with us a wonderful friendship even at times when we least deserve it, we couldn't be more lucky to have you! We know we can never thank you enough. To our advisers, for sharing all the knowledge we need, for the time, patience and effort, and for simply believing that we can, thank you Sir and Ma'ams. You being with us every step of the way made it all easier to take. And lastly, but most importantly, to God, our Creator, for this awesome life and for giving us these awesome people, thank you! All our success we offer to you and only you Lord. Japs, Juni and Alyssa

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TABLE OF CONTENTS Page Title Page ..... Abstract Acknowledgement ... Table of Contents . List of Figures .. i ii iii iv vi

List of Tables ... vii List of Appendices ... viii

CHAPTER 1 THE PROBLEM AND ITS SCOPE INTRODUCTION . Background of the Study Objectives of the Study .. Hypotheses . Significance of the Study ... Scope and Limitations of the Study ... Definition of Terms 2 REVIEW OF RELATED LITERATURE Lead Removal of Heavy Metal in Contaminated Water . Phytotechnology . The Duckweeds .. Species Identification . Duckweed Composition . Heavy Metal Accumulation by Duckweeds ... Biosorption of Duckweeds . 5 5 6 7 7 8 9 9 1 1 3 3 3 4 4

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MATERIALS AND METHODS Collection and Preparation of Research Materials and Equipment . 10 Research Procedure/Experimental Design . 11 Gathering of Data ... 13 Treatment of Data ... 24 Ways of Proper Disposal 24

RESULTS AND DISCUSSIONS Findings .. 25

CONCLUSION AND RECOMMENDATIONS ... 29

APPENDICES .. 30 REFERENCES . 38

LIST OF FIGURES Figure 1 The flow of processes on the determination of the effect of pH and agitation time on lead biosorption efficiency on aqueous solution by duckweed (Lemna sp.).. 14 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 The UPV Facultative pond covered with duckweeds (Lemna sp.) ..15 Collection of samples using a long-handed mesh sampler ......... 15 Collected duckweed samples with impurities . 16 Washing of collected samples .... 16 Clean duckweed samples .... 17 Air-drying of samples . 17 Air-drying of samples . 18 Cabinet-drying of samples ... 18 Dried duckweeds 19 Weighing of dried duckweeds .... 19 Preparation of Lead solution .. 20 Prepared Lead solution ... 20 pH adjustment . 21 Moisture analysis .... 21 The experimental setup ... 22 Filtration of mixture .... 22 Bottled filtrate sample . 23 Packed filtrate samples ready for analysis .. 23 A pH vs. % Removal plot for 45-minute agitation time ..... 26 A pH vs. % Removal plot for the 90-minute agitation time ... 26 A combined pH vs. % Removal plots of the 45 and 90-minute agitation time ... 27 23 Plots of Agitation Time vs. % Removal for different pH levels . 28 Page

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LIST OF TABLES Table 1 2 Page Specifications of treatments used in the study ... 12 Summary of results on the effect of variations in pH and agitation time on lead biosorption efficiency from aqueous solution by duckweed (Lemna sp.) . 25

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LIST OF APPENDICES Appendix 1 The Research Paradigm on the Effect of pH and Agitation Time on Lead Biosorption Efficiency on Aqueous Solution by Duckweed (Lemna sp.) ... 31 2 3 4 Lead Solution Preparation (Serial Dilution) .. 32 . Storage and Preservation of Samples . 33 Certificate of Analysis from the Department of Science and Technology Regional Office No. 7 (DOST VII) . 34 5 6 7 Calculations for % Removal of Lead 35 Raw Data for Moisture Content Analysis ... 36 Letter to the Chancellor for Permission to have Access to the UPV Facultative Pond ..37 Page

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CHAPTER 1

INTRODUCTION

Background of the Study With the growing awareness on environmental issues, global interest nowadays is on the development of the cost effective and environment friendly technologies for the remediation of soils and wastewaters polluted with toxic substances including heavy metals. Occurrence of toxic metals in bodies of water affects the lives of people within its locality because of their dependence upon these water sources for their daily requirements (Rai et al., 2002). Aquatic food contaminated with toxic metals may be consumed through its introduction in the food chain and may cause serious health hazards. Among all metals, the public is at greater risk from lead than any other metal (Scott, 1995). In the tropical region of Asia, environmental pollution is of a greater threat due to lack of wastewater treatment facilities as it is mostly occupied by low to middle income countries with rapid industrial and urban development, and most wastewater treatment technologies, like chemical precipitation and coagulation, are known to be costly for municipalities and small scale polluting industries (Trihadiningrum, Verheyen and De Pauw, 1997). In the Philippines for example, despite the improving household garbage collection, still 90 percent of the sewage is not treated and disposed in an environmentally sound manner. Only 10 percent of the countrys total population is connected to sewers and others rarely maintain adequate on-site sanitation. The increase in water pollution which is caused primarily by fragmented water management, weak enforcement of regulations, and poor planning are preventing adequate responses, is degrading the countrys groundwater, rivers, lakes, and coastal areas and the quality of pollution is estimated to be at US$1.3 billion, including health costs, losses in fisheries production, and impact on tourism (World Bank, 2011). The good thing however is that tropical regions are characterized by whole year of solar energy resource which proves to be beneficial for supporting plant based
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wastewater treatment technology. Technologies that are of this type is known as costeffective and publicly accepted. One example of these said technologies which already receives increasing appraisal nowadays is phytotechnology. Phytotechnology is the application of science and engineering in resolving environmental issues with the use of plants. This implies a wider understanding of the importance of plants and their beneficial role in the society and in natural systems (Mangkoedihardjo, 2007). Phytotechnology is not only known as cost-effective but it as well offers aesthetics and supports wild life habitat (USEPA, 1993). Biosorption is a new cost-effective phytotechnology that is of significant use today. It is a process that involves a solid phase (sorbent or biosorbent) and a liquid phase (solvent, usually water). The mechanism of metal biosorption is a complicated process which would either involve a living or a non-living biomass (Das, R., & Karthika, 2007). Plants such as algae and other macrophytes are known to be good biosorbents for biosorption as they tend to concentrate metals to exceptionally high levels and may therefore help in reclamation of effluents containing heavy metals. Duckweeds are widely used macrophyte for this purpose as they have a wide range of temperature tolerance, are easily harvested and are capable of rapid growth. Duckweeds can tolerate and accumulate high concentrations of heavy metals and organic compounds (Ornon et al., 1984). Various studies abroad had also been conducted to show that duckweeds can be an effective medium to clean waste water contaminated with heavy metals.

Objectives of the Study The study aimed to determine the effect of variations in conditions in the biosorption efficiency of duckweed (Lemna sp.) in lead removal. Specifically, the study 1. determined the effect of pH 2. and agitation time on the efficiency of duckweed (Lemna sp.) as bioadsorbent in terms of % removal of lead from the aqueous solution.

Hypotheses of the Study The following hypotheses were put forward: 1. There exists no relationship between the % removal of lead by duckweed (Lemna sp.) and pH. 2. There exists no relationship between the % removal of lead by duckweed (Lemna sp.) and agitation time.

Significance of the Study Heavy metal pollution is a pressing problem the world faces today. As compared to most organic materials, heavy metals cannot be transformed by organisms, resulting to their accumulation in water, soil, bottom sediments and living organisms. These pollutants usually result from sewage works, industrial processing and irrigation run-off. Most of the heavy metals are toxic or carcinogenic in nature and pose a threat to human health and the environment especially our waters. In view of the fact that theres an undeniable need for water purification and reuse, duckweeds potential in removing heavy metals is a promising solution. In addition to that, duckweeds can be used as indicator species as their content of heavy metals can be used to indicate potential pollution levels of waters. In the Philippines, there are limited studies on the use of duckweeds as biological means of removing heavy metals from waste water. Most of the literatures available were conducted outside the country. Therefore, the study will be significant for future researches about duckweeds because of its local setting. The result of the study on the
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efficiency of biosorption of duckweed can be used as basis for industrial waste water treatment plant with heavy metal effluents.

Scope and Limitations of the Study This study focused only on the investigation of the effect of pH and agitation time on the efficiency of duckweed (to be harvested from UPV Facultative Pond) in lead removal from aqueous solution through biosorption. It was conducted at the Laboratory of the School of Technology, Miag-ao, Iloilo during the First Semester of Academic Year 2011-2012. The variable evaluated was the biosorption efficiency of duckweed in terms of % removal of lead. Also, the effect of agitation time and the pH of the solution were given specific attention.

Definition of Terms Biosorption. Removal of heavy metals on polluted water by non-living, inactive biomass. Blank. A portion of the prepared lead solution. Digestion. Use of acid and heat to break organo-metallic bonds and free ions for analysis. Duckweed. One of the smallest, free-floating aquatic macrophytes that take up metals in water. Lead. A toxic heavy metal usually present in industrial effluents. Percent Removal. The initial concentration of lead minus the final concentration over the initial concentration in the aqueous solution multiplied to 100%. Biosorption Efficiency. The efficiency of the biosorption process in removal of lead.

CHAPTER 2 REVIEW OF RELATED LITERATURE

Technological advancement has given an increasing problem of heavy metals which are listed as priority pollutants by the US Environmental Protection Agency (Rai,P.K,2007). Soil and water pollution by heavy metals is a serious environmental problem worldwide. Unlike organic pollutants, they cannot be degraded by microorganisms. Heavy metals can make its way to the food chain and be consumed by people. Different metals have wide range of applications. Even though a metal doesnt have a role in the manufacturing process, we can still be exposed to it. Considering levels of exposure and levels necessary to produce toxic effects, the public is at greater risk from lead than any other metal (Scott, 1995).

Lead Lead is used in storage batteries. Lead alloyed with antimony and tin is used as a sheath to pigments, in solders and in ammunition. Before, leaded gasoline was used as an anti-knock additive. But because of its environmental impact, it was changed to unleaded. The toxic effects of lead on the body are varied but greatly affect the nervous system. Lead poisoning symptoms include convulsions, hallucinations, coma, weakness and tremors. It can also lead to anemia and kidney damage. Once exposed to lead, it is incorporated into the bones and is released very slowly (Scott, 1995).

Removal of Heavy Metal in Contaminated Water The removal of heavy metals from industrial wastewater has been the focus of some studies. To reduce the heavy metals concentration in water, conventional methods such as chemical precipitation, ion exchange, reverse osmosis, and adsorption have been proposed. These processes either have low efficiencies or extremely expensive. Some studies examine the potential of natural materials in removing heavy metals. Natural materials that are available in large quantities or certain waste from agricultural processes

may have the potential to be used in the removal. In addition, they represent unused resources, widely available and are environmentally friendly (Afkhami et al., 2008).

Phytotechnology Phytotechnology describes the application of science and engineering in examining environmental problems with the use of plants. In phytotechnology, the plants used serve as pump and treat system as they take up water soluble contaminants through their roots and transport them through various plant tissues, where they can be metabolized or volatilized (Doty et al., 2007). Terrestrial and aquatic plants have the ability to acquire and accumulate metal ions such as Pb, Cu, Mn and Zn. Some metal ions are essential for plant growth and development. Thus, these plants can be used to remove the heavy metals from contaminated sites. Because of their ability to grow under polluted conditions, some aquatic plants have been investigated for their potential to remove the heavy metals and improve the quality of the wastewater (Reiner et al., 1993). These freshwater vascular plants when combined with macroscopic algae are collectively known as macrophytes. According to Outridge and Noller (1991), heavy metal uptake and retention by the macrophytes are controlled by the sediment geochemistry, water physicochemistry, plant physiology and genotypic differences. Sediment geochemistry and water

physicochemistry control metal speciation in sediments and water. Plant physiology and genotypic differences control the ability of plants to accumulate plant- available forms of the metal. Freshwater vascular plants exhibit several advantages when compared to algae. These plants have longer life cycles. They have a higher degree of tolerance to most heavy metal that are causing pollution. Availability of a large sample for analysis is possible because of larger biomass compared to algae (Brooks and Robinson). Remediation of polluted water by these plants may be achieved in two ways. The first uses the monospecific pond cultures of free-floating plants. Free floating macrophytes are generally restricted to sheltered habitats and slow-flowing waters. Their nutrient absorption is completely from the water, and most of these macrophytes are found in water bodies rich in dissolved salts. The plants accumulate the pollutants until
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an equilibrium condition is achieved. Then they are removed from the pond. Problems that can be encountered in this method are the proper disposal of the plant contaminated with heavy metals after they are removed from the pond. Also, unwanted pathogens may destroy the whole of a monospecific culture. Then, there is also the need for continuous harvesting that requires specialized equipment second method involves growing rooted emergent in trickling bed filters. Rhizophore microbes are responsible for the uptake of trace elements in these systems (Brooks and Robinson). Aquatic plants, such as water hyacinth (Eichhorniacrassipes), pennywort (Hydrocotyle umbellate), small water fern (Azolla sp.), water lettuce (Pistia sp.) and duckweeds are known to be effective in single pass wastewater treatment processes (Mishra,Tripathi, 2009).

The Duckweeds Attention given on the duckweed has been due to its wide range of applications or possible ways it can be utilized. El-Shafai et al, in 2006 studied the uptake of nutrients and mineral contaminants from wastewater effluent. Duckweed can also be used as animal feed (Cheng and Stomp, 2009). Iqbal in 1999 studied the potential of duckweed as compost. Recent studies utilize duckweed in bio-energy production (Fedler et al, 2007; Cheng and Stomp, 2009). Duckweed is one of the smallest macrophytes, free-floating aquatic plants found distributed worldwide. They are monocotyledons belonging to the family Lemnaceae. Duckweeds belong to five genera; Lemna, Landolita, Spirodela, Wolfia and Wolffiella. About 40 species are known worldwide. Duckweeds have flattened minute, leaf-like oval to round fronds from about 1 mm to less than 1cm across. The bulk of the frond is composed of chlorenchymatous cells separated by large intracellular spaces that are filled with air or other gases which provides buoyancy to the plant.

Species Identification In open water, some species develop root-like structures to stabilize the plant or to help in obtaining nutrients in dilute concentrations. The fronds of Spirodela and
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Lemnaare flat, oval and leaf like. On each frond of the Spirodela species, two or more thread-like roots are present while Lemna has only one. Wolfia fronds are usually sickle shaped. Wolffiella is boat shaped Wolffiellaand Wolfia are thalloid and have no roots. They are also much smaller compared to Spirodela or Lemna. Species of the genus Spirodela have the largest fronds measuring to about 20 mm across. The size of the fronds of Lemna species typically average between 6-8 inches while those of the Wolffia species are about 2 mm or less in diameter. In all genera, each mother frond produces a considerable number of daughter fronds during its lifetime. It can produce as many as 20 daughter fronds during its lifetime, which lasts for a period of 10 days to several weeks. The daughter fronds are produced alternatively and in a pattern. In Spirodela and Lemna, daughter fronds appear in a pattern from two pockets which is close to where the roots arise in each side of the mature frond. In Wolffiella and Wolfiaonly one pocket exists. Some cells of Lemna and Spirodela have needle like raphides which are presumably composed of calcium oxalate. The upper epidermis in the Lemna is highly cutinized and is unwettable. Stomata are on the upper side in all the genera. Roots in both Spirodela and Lemna are adventitious. The plant reproduces both vegetatively and sexually. The flowering occurs sporadically and unpredictably. Turion is a starchy survival frond formed by many species of duckweeds to survive at low temperature. In cold weather, it sunk at the bottom of the pond where it remains dormant until warm water triggers the resumption of normal growth.

Duckweed Composition Duckweed is composed of water, mineral elements, and organic matter. The composition of the duckweeds varies per species. It has been reported on a study by Cross in 2006 that the freshwater duckweed fronds water content ranges from 87% to 97% depending on the species. Chemical analyses shows varying composition of crude protein, ash, fibre and water content, fat and mineral content depending on the species, the harvest location and the water source. The crude protein content of duckweeds depends mostly on the nitrogen content of the water upon which they grow. Cheng and
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Stomp (2009) reported starch content values ranging from 3- 75% of the dry weight depending on the species of duckweeds. Accumulation of starch in the plant biomass is possible during periods of dormancy which can be achieved by varying growth conditions like pH, nutrient concentration, and temperature (Cheng and Stomp, 2009).

Heavy Metal Accumulation by Duckweeds All members of the duckweed family can tolerate and accumulate high concentrations of heavy metals and organic compounds from sources. Many reports are available on the uptake of metal ions by duckweeds and the numerous interactions that occur. Duckweeds will uptake and concentrate Cd, N, Cr, Zn, Sr, Co, Fe, Mn, Cu, Pb, Al and even Au.

Biosorption of Duckweeds Biosorption is a wastewater remediation approach in which removal of heavy metals from aqueous solutions by non-live, inactive biomass. The non-live biomass used is called biosorbent. Some biosorbent show high heavy metal binding capacity. There are various types of biomass, such as leaves, wood or agricultural residues. Jameel M. Dhabab in January 2011 studied the removal of some heavy metal ions from their aqueous solutions by duckweeds using biosorption technique. The result showed that removal percentage was 94% for Pb2+, 72% for Zn2+, 65% for Cu2+, and 50% for Fe2+. The efficiency of adsorption depends on pH value; it increases with the increase of pH value at 2 to 8 in range time between 60 to 90 min. The most percentage of removal in the main way of loading weigh was 1.5 grams and 50 ml mixed metal ions solution at 50 ppm concentration of each metal. The duckweed is found to be a promising adsorbent for the removal of metal cations from mixed metal ions solution, representing an effective and environmentally clean waste matter.

CHAPTER 3

MATERIALS AND METHODS

The flow of processes on the determination of the effect of pH and agitation time on lead biosorption efficiency on aqueous solution by duckweed (Lemna sp.) is shown in Figure 1, the Schematic Diagram.

Collection and Preparation of Research Materials and Equipment Preparation of Duckweed Samples. The researchers collected about two (2) kilograms of fresh duckweeds (Lemna sp.) from the surface of waste water pond (UPV Facultative Pond) (see Figures 2 and 3) using a long-handed mesh sampler. The samples then were brought to the School of Technology Laboratory, UPV, Miag-ao, Iloilo. The gathered samples (see Figure 4) were washed with tap water to partially clean the plant surface (see Figure 5). Foreign materials from the pond that mixed up with the samples upon gathering were removed manually. To ensure the removal of other impurities, distilled water was used for rinsing the material. Then, the clean samples (see Figure 6) were air-dried for twenty-four (24) hours (see Figures 7 and 8) and cabinetdried for three (3) hours at fifty (50) to sixty (60) C (see Figure 9). Dried duckweed sample (see Figure 10) was placed in clean plastic bag and weighed (see Figure 11). Final moisture contents before and after cabinet-drying were monitored (see Figure 15). Preparation of Lead Solution. Six (6) L of about fifty (50) mg/L (ppm) lead solution was prepared from Lead Nitrate (Pb(NO3)2) salt using distilled water as the diluent (see Figures 12 and 13, also see Appendix 3). The final concentration of the prepared solution was determined by subjecting a sample of the solution to lead analysis using Atomic Absorption Spectrometer (AAS). Five hundred (500) mL of the solution was used for six (6) treatments. The pH levels were varied for each treatment. The pH adjustment was carried out using 0.1 M Hydrochloric Acid (HCl) or 0.1 M Sodium Hydroxide (NaOH) (see Figure 14). Two (2)

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sets of samples were prepared making twelve (12) in total (see Table 1. Specifications of treatments used in the study). Preparation of Sample Containers. Unused five hundred (500) mL Polyethylene (PET) bottles were prepared as sample containers prior to the experiment. The bottles were ensured to be clean before use.

Research Procedure/Experimental Design Moisture Content Analysis. One (1) gram of duckweed sample was analysed for moisture content using a moisture analyser (see Figure 14). Biosorption Experiment. The experiment was carried out in one (1) L glass beakers containing five hundred (500) mL of the prepared lead solution of known pH. For each treatment, seven (7) grams of the duckweeds (wet basis) samples was introduced into the solution. To maximize the contact of the plant material with the solution, Phipps and Bird Mechanical Stirrer was used to agitate the system at one hundred forty (140) rotations per minute (rpm) were used in the study. The first set of samples was subjected to forty-five (45) minute agitation. The sample procedure was repeated on the other set of treatments agitated for ninety (90) minutes (see Figure 16). The mixtures were filtered through a filter paper (see Figure 17). Four hundred (400) mL of the filtrate were stored in clean sample containers and sealed prior to sample digestion and analysis (see Figures 18 and 19). The specification of the treatments used in the study is shown in Table 1.

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Table 1. Specifications of treatments used in the study. Treatments pH Mass of Dried Duckweeds Initial Lead Concentration

45-minute Agitation TA,45 TB,45 TC,45 TD,45 TE,45 Blank TA,90 TB,90 TC,90 TD,90 TE,90 Blank 4.0 6.0 7.0 8.0 10.0 Unaltered (5.0) 4.0 6.0 7.0 8.0 10.0 Unaltered (5.0) 0.0 g 7.0 g ~50 mg/L 0.0 g 90-minute Agitation 7.0 g ~50 mg/L

Table 1 shows the specification of treatments used in the study. The lead solutions for Treatments A to E were adjusted to pH values of 4.0, 6.0, 7.0, 8.0 and 10.0 respectively; with Treatment C at pH 7.0 as the control; as it is neither acidic nor basic. pH values were chosen in a way that with enough number of treatments, it would produce a clear result of how acidity or basicity affects the adsorption capacity of duckweed. All the treatments were then subjected to forty-five (45) and ninety (90) minutes agitation along with the blanks having no duckweed content.

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Gathering of Data Moisture Content Analysis. The analyses were repeated until three trials (% moisture) are obtained that pass the Q-test. Lead Analysis. The samples were brought to the Department of Science (DOST) and Technology Regional Office No. 7 Regional Standards and Testing Laboratory (Laboratory No. 11-09-154-03) for the analysis using Atomic Absorption Spectrometer. Samples were digested using Nitric acid (HNO3) and diluted to volume prior to the analysis.

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Duckweeds Harvesting Cleaning/Washing Distilled Water Pb2+ Air Drying Cabinet Drying Weighing pH Adjustment Moisture Content Analysis Blank TA TB TC TD TE Moisture Content

Stirring

45 minutes

90 minutes

Filtration Packing/Storing Sample Digestion Pb Analysis by AAS Pb Content % Pb Removal Figure 1. The flow of processes on the determination of the effect of pH and agitation time on lead biosorption efficiency on aqueous solution by duckweed (Lemna sp.)
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THE EFFECT OF PH AND AGITATION TIME ON LEAD BIOSORPTION EFFICIENCY FROM AQUEOUS SOLUTION BY DUCKWEED (Lemna sp.)

Figure 2. The UPV Facultative pond covered with duckweeds (Lemna sp.).

THE EFFECT OF PH AND AGITATION TIME ON LEAD BIOSORPTION EFFICIENCY FROM AQUEOUS SOLUTION BY DUCKWEED (Lemna sp.)

Figure 3. Collection of samples using a long-handed mesh sampler.

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THE EFFECT OF PH AND AGITATION TIME ON LEAD BIOSORPTION EFFICIENCY FROM AQUEOUS SOLUTION BY DUCKWEED (Lemna sp.)

Figure 4. Collected duckweed samples with impurities.

THE EFFECT OF PH AND AGITATION TIME ON LEAD BIOSORPTION EFFICIENCY FROM AQUEOUS SOLUTION BY DUCKWEED (Lemna sp.)

Figure 5. Washing of collected samples.

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THE EFFECT OF PH AND AGITATION TIME ON LEAD BIOSORPTION EFFICIENCY FROM AQUEOUS SOLUTION BY DUCKWEED (Lemna sp.)

Figure 6. Clean duckweed samples.

THE EFFECT OF PH AND AGITATION TIME ON LEAD BIOSORPTION EFFICIENCY FROM AQUEOUS SOLUTION BY DUCKWEED (Lemna sp.)

Figure 7. Air-drying of samples.

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THE EFFECT OF PH AND AGITATION TIME ON LEAD BIOSORPTION EFFICIENCY FROM AQUEOUS SOLUTION BY DUCKWEED (Lemna sp.)

Figure 8. Air-drying of samples.

THE EFFECT OF PH AND AGITATION TIME ON LEAD BIOSORPTION EFFICIENCY FROM AQUEOUS SOLUTION BY DUCKWEED (Lemna sp.)

Figure 9. Cabinet-drying of samples.

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THE EFFECT OF PH AND AGITATION TIME ON LEAD BIOSORPTION EFFICIENCY FROM AQUEOUS SOLUTION BY DUCKWEED (Lemna sp.)

Figure 10. Dried duckweeds.

THE EFFECT OF PH AND AGITATION TIME ON LEAD BIOSORPTION EFFICIENCY FROM AQUEOUS SOLUTION BY DUCKWEED (Lemna sp.)

Figure 11. Weighing of dried duckweeds.

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THE EFFECT OF PH AND AGITATION TIME ON LEAD BIOSORPTION EFFICIENCY FROM AQUEOUS SOLUTION BY DUCKWEED (Lemna sp.)

Figure 12. Preparation of Lead solution.

THE EFFECT OF PH AND AGITATION TIME ON LEAD BIOSORPTION EFFICIENCY FROM AQUEOUS SOLUTION BY DUCKWEED (Lemna sp.)

Figure 13. Prepared Lead solution.

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THE EFFECT OF PH AND AGITATION TIME ON LEAD BIOSORPTION EFFICIENCY FROM AQUEOUS SOLUTION BY DUCKWEED (Lemna sp.)

Figure 14. pH adjustment.

THE EFFECT OF PH AND AGITATION TIME ON LEAD BIOSORPTION EFFICIENCY FROM AQUEOUS SOLUTION BY DUCKWEED (Lemna sp.)

Figure 15. Moisture analysis of dried duckweeds.

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THE EFFECT OF PH AND AGITATION TIME ON LEAD BIOSORPTION EFFICIENCY FROM AQUEOUS SOLUTION BY DUCKWEED (Lemna sp.)

Figure 16. The experimental setup.

THE EFFECT OF PH AND AGITATION TIME ON LEAD BIOSORPTION EFFICIENCY FROM AQUEOUS SOLUTION BY DUCKWEED (Lemna sp.)

Figure 17. Filtration of mixture.

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THE EFFECT OF PH AND AGITATION TIME ON LEAD BIOSORPTION EFFICIENCY FROM AQUEOUS SOLUTION BY DUCKWEED (Lemna sp.)

Figure 18. Bottled filtrate samples.

THE EFFECT OF PH AND AGITATION TIME ON LEAD BIOSORPTION EFFICIENCY FROM AQUEOUS SOLUTION BY DUCKWEED (Lemna sp.)

Figure 19. Packed filtrate samples ready for analysis.

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Treatment of Data Average Moisture Content (AMC). The data (% moisture) at different trials were averaged.

Lead Concentration. The lead concentrations for each treatment as analysed was plotted against pH and agitation the time and interpreted. Percent Removal. The efficiency of biosorption was assessed in terms % removal of lead from the aqueous solution. % removal by duckweed was calculated using the following equation:

Where

and

were the initial and final concentration (ppm) of lead in the

solution respectively. The percent efficiencies were plotted against pH.

Ways of Proper Disposal The remaining solutions containing lead were contained in reagent bottles. The duckweeds residue exposed to lead were sealed in bottles and were disposed in a specific area in the UPV dumpsite reserved for waste chemicals.

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CHAPTER 4

RESULTS AND DISCUSSIONS Findings Cabinet-drying of duckweeds resulted to reduction of moisture from 87.65 % to 47.35 % (see Appendix 6). The final concentration of the prepared lead solution as determined by AAS was found to be forty-eight (48.0) ppm (see Appendix 4). This concentration served as the initial concentration (Ca) for all treatments. The results of the experimentations on the determination of the effect of pH and agitation time on lead biosorption efficiency on aqueous solution by duckweed (Lemna sp.) are summarized in Table 2.

Table 2. Summary of results on the effect of variations in pH and agitation time on lead biosorption efficiency from aqueous solution by duckweed (Lemna sp.). Treatments A B C D E Blank pH 4.0 6.0 7.0 8.0 10.0 5.0* 45-minute Agitation Cf a (ppm) 48.0 45.9 42.1 21.4 9.76 45.3 % removalb 0.0 4.38 12.3 55.4 79.7 5.63 90-minute Agitation Cf a (ppm) 45.8 44.7 42.7 16.1 9.64 43.0 % removalb 4.58 6.88 11.04 66.5 79.9 10.4

Note: a - see Appendix 4; b - see Appendix 5; * - unaltered Given the initial and final lead concentrations of the treatments, the percentage of removal of lead from the aqueous solution was calculated and tabulated in Table 2. The calculated percentage of removal illustrates the efficiency of duckweed in removing lead from the solution. As can be observed in Table 2, the combined effect of increasing the pH and agitation time produced a corresponding increase in the removal of lead.

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The positive results for the removal of lead rendered by two blank samples imply the possibility of the effect of agitation alone on the removal of lead. However, the factors that can be attributed to these results are no longer within the scope of this study. Giving specific attention for each agitation time, the plot of pH vs. % removal for the 45-minute agitation as shown in Figure 20, illustrates clearly that as pH increases, the amount of lead removed from the solution also increases. Same observation can be deduced for the 90-minute agitation time as shown in Figure 21.

90
80 70 % Removal 60 50 40 30 20 10 0 0 2 4 6 pH 8 10 12

Figure 20. A pH vs. % Removal plot for 45-minute agitation time.

90 80 70 % Removal 60 50 40 30 20 10 0 0 2 4 6 pH 8 10 12

Figure 21. A pH vs. % Removal plot for 90-minute agitation time.

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90
80 70 % Removal 60 50 40 30 45 90

20
10 0 0 2 4 6 pH 8 10 12

Figure 22. A combined pH vs. % Removal plots of the 45 and 90-minute agitation time.

As can be seen in Figure 22, the 90-minute agitation time exceeds the 45-minute agitation time in terms of % removal. Although there is an overlap in the pH value 7.0 for both 45 and 90-minute agitation, in general, 90-minute agitation time has higher % removal. This is because, as the agitation time increases, there is an increased possibility of more contact between the duckweed and the solution. So there is more chance that the duckweed can increase its uptake of lead.

27

90 80 70 60 % Removal 50 pH 4 pH 6 pH 7

40
30 20 10 0 0 20 40 60 80 100 Agitation Time

pH 8
pH 10

Figure 23. Plots of Agitation Time vs. % Removal for different pH levels.

For Figure 23, at pH 4.0 and 45-minute agitation time, there was no lead removal. This is because the contact between the duckweed and the solution is not yet enough for removal to take place but for 90-minute agitation, some amount of lead was already removed from the solution. The % removal was higher for both 45 and 90-minute agitation time at pH 10.0. In low pH values, binding sites are generally protonated or positively charged by the hydronium ions present. Thus, repulsion occurs between the metal cation (Pb2+) and the adsorbent. At a higher pH values, binding sites start deprotonating, hydroxide ions prevail, and makes different function groups available for metal binding. In general, cation binding increases as pH increases (Forsner and Wittman, 1981; Esposito et al., 2002).

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CHAPTER 5

CONCLUSION AND RECOMMENDATIONS

Conclusion The outcome of the study has generally shown the great potential of duckweed in removing heavy metals specifically lead and that varying conditions of the process could significantly magnify the results. Based on the results obtained after the experiment, there exist a relationship between the percentage of removal of lead from aqueous solution by duckweed and both varying pH and agitation time. As it has been shown, increasing pH value and agitation time produces a greater yield of lead removed from the solution. The pH value of 10.0 at 90-minute agitation gave the highest amount of lead removed.

Recommendations In view of the results and the conclusion, the researchers recommend that further study must be made on the effect of varying the process conditions (i.e. concentration of solution, pH, agitation time, mechanical stirring etc.) on the heavy metal biosorption efficiency of duckweed. It is recommended as well that further studies must be conducted to determine the optimum condition that would give optimum yield of removal. Also, particular attention must be given to the adsorption isothermal studies of duckweed. In addition to that, the researchers also suggest that thorough investigation must be done regarding the efficiency of duckweed to remove other heavy metals like chromium, zinc and iron and the kinetics behind it.

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APPENDICES

30

Appendix 1. The Research Paradigm on the Effect of pH and Agitation Time on Lead Biosorption Efficiency on Aqueous Solution by Duckweed (Lemna sp.)

Independent Variables

Dependent Variable

Treatment A pH 4.0, 7.0 g dried duckweed, ~50 ppm Lead, 45 and 90-minute agitation

Treatment B pH 6.0, 7.0 g dried duckweed, ~50 ppm Lead, 45 and 90-minute agitation

Treatment C (Control) pH 7.0, 7.0 g dried duckweed, ~50 ppm Lead, 45 and 90-minute agitation % Removal Treatment D pH 8.0, 7.0 g dried duckweed, ~50 ppm Lead, 45 and 90-minute agitation

Treatment E pH 10.0, 7.0 g dried duckweed, ~50 ppm Lead, 45 and 90-minute agitation

Blank pH 5.0, 0.0 g dried duckweed, ~50 ppm Lead, 45 and 90-minute agitation

31

Appendix 2. Lead Solution Preparation (Serial Dilution) The Lead solution was prepared as followed, 1. 1.6135 grams Lead Nitrate (Pb(NO3)2) salt was weighed in an analytical balance. 2. The salt was dissolved with 50 mL distilled water in a 100 mL beaker. 3. The solution was decanted into a 1000 mL volumetric flask. 4. The beaker was washed thrice with distilled water pouring the washings into the flask. 5. The flask was filled up to mark with distilled water. 6. 600 mL aliquot of the prepared solution was transferred into a larger container container and diluted with 5.4 L distilled water.

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Appendix 3. Storage and Preservation of Samples

Samples were stored in clean polyethylene (PET) bottles away from any potentially contaminating source prior to analysis. Since the analysis was not done immediately, the samples were refrigerated in order to minimize microbial activity. An additional advantage of refrigeration as a preservation method is that it neither affects the sample composition nor interferes with any analytical method. (Nollet, 2007).

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Appendix 4. Certificate of Analysis from the Department of Science and Technology Regional Office No. 7 (DOST VII) Regional Standards and Testing Laboratory

34

Appendix 5. Calculations for % Removal of Lead

Sample calculations: For TA (4.0 pH, 45-minute agitation):

For TA (4.0 pH, 90-minute agitation):

Summary of the results of % Removal calculation of each treatment for both agitation times. % Removal Treatments pH 45-minute Agitation 90-minute Agitation A 4.0 0.0 4.58 B C D E BL 6.0 7.0 8.0 10.0 5.0 4.38 12.3 55.4 79.7 5.63 6.88 11.04 66.5 79.9 10.4

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Appendix 6. Raw Data for Moisture Content Analysis Moisture Content (%) Air-dried Duckweeds Cabinet-dried Duckweeds Q-test:
| |

Trials 1 87.34 50.40 2 86.40 46.97 3 89.21 44.69

; if

, the suspected value is accepted. At 90% degrees

of freedom, Qtab = 0.941

Moisture Content (%) 87.34 86.40 89.21

| | | | | |

Qcal
| | | | | |

Result Accept Accept Accept

Average Moisture Content of air-dried duckweeds: 87.65 %

Moisture Content (%) 50.4 46.97 44.69

| | | | | |

Qcal
| | | | | |

Result Accept Accept Accept

Average Moisture Content of cabinet-dried duckweeds: 47.3533 %

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Appendix 7. . Letter to the Chancellor for Permission to have Access to the UPV Facultative Pond

37

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