Utilization of Sunflower (HELIANTHUS ANNUUS L.

) for the
absorption of Lead in Contaminated Soil

A Research submitted to

Malayan High School of Science

Paz Guazon Street, Pandacan, Manila

In partial fulfillment of the requirements in

Practical Research II

Narag, Roy Nicolo

Navalta, Ivan Christopher

Simbulan, Ron Jerome

Tiu, Immanuel

Andrade, Vince

Salanguit, Wally

Introduction
Background of The Study

Among heavy metal, lead provides numerous application possible with its chemical

properties (Karrari el al, 2012). With its diverse properties, lead is used for many industries,

including mining, smelting, refining, battery manufacturing and others (Mohammadi et al, 2008).

However, due to its widespread usage, lead has become rampant in the environment, which has

become a major concern for public health. Medicine history has also dictated lead poisoning to

be a well-known disease.

With lead being widespread in the environment, it raised concern that land, water and air

are major sources of lead. Exposure to lead for general population mainly from airborne dusts

containing lead particles and from food or water contaminated by lead. 15-30% of lead in the

environment is inhaled, while 70-86% of it is ingested (Kalantri et al, 2009). Their pollution in

water and soil is giving a threat to the humans, plants and animals.

Phytoremediation is the direct use of living green plants for in situ, or in place, removal,

degradation, or containment of contaminants in soils, sludges, sediments, surface water and

groundwater (http://www.unep.or.jp). Plants are an extraordinary organism equipped with

astounding metabolic and absorption capabilities. Phytoremediation deals with the growing of

plants in a contaminated matrix, for a required growth period, to remove contaminants from the

matrix, or facilitate immobilization (binding/containment) or degradation (detoxification) of the

pollutants (http://www.unep.or.jp). The plants can be harvested at a later date, processed and

disposed. Various types of vegetation including trees, grasses, aquatic plants, are used for

phytoremediation.
 In the Philippines, certain areas like western Luzon is known for soils rich in heavy

metals.( http://www.greenpeace.org). Metal-absorbing plants are of significant interest for

environmental advocates. Some plants can be used to remove dangerous metallic materials from

a polluted ecosystem. Once the plant has absorbed a significant amount of metal, they can also

be harvested for their commercially valuable contents.

Objectives

- To know if sunflower can be used for absorbing heavy metals such as lead and to know

how much lead it can absorb.

- To know if, via phytoremediation, sunflower could be a viable plant to use for filtering

the soil/water.

Significance of the Study

This research is significant to all the people in the society, especially to those who live in places

where soil contamination/pollution is very common. Utilization of plants that can absorb heavy

metals like lead would be a vital key for people to solve the contamination issues.

This research is also significant to scientists and researchers that are currently studying about the

possible ways in effectively removing heavy metals in soil with the use of plant, since this

research indicates if whether or not sunflower will have a potential of absorbing heavy metal

such as lead.

Hypothesis

Ho The plant sunflower can be used for phytoremediation of lead contaminated soil.
 Ha The plant sunflower cannot be used for phytoremediation of lead contaminated soil.

Scope and Limitation

The researchers will determine the effectiveness of sunflower in absorbing heavy metals

specifically lead only. Therefore, this research only answers the question if whether or not

sunflower can be used for phytoremediation of lead contaminated soil.

Definition of Terms

Phytoremediation - Phytoremediation is a bioremediation process that uses various types of

plants to remove, transfer, stabilize, and/or destroy contaminants in the soil and groundwater.

There are several different types of phytoremediation mechanisms (http://www.cpeo.org).

Various types of vegetation including trees, grasses, aquatic plants, are used for

phytoremediation.

Sunflower - The sunflower, or Helianthus annuus, is an annual flower that is famous for its

extremely large and vividly yellow blooms. These plants are native to North America, but they

are widely cultivated all over the world today as ornamentals, food plants and for their valuable

oil. It is also one of the many plants that are now known to aid in phytoremediation, a process

that employs various types of plants to remove, transfer, stabilize, and/or destroy contaminants in

the soil, water and air (http://farmersalmanac.com).

Lead - Lead is a naturally occurring toxic metal found in the Earths crust. Its widespread use

has resulted in extensive environmental contamination, human exposure and significant public

health problems in many parts of the world.

Review of Related Literature

Sunflower

It is determined that the sunflower was domesticated around 1000 BCE in North America,

presumably for the motive of using the oil-rich and highly nutritious seeds. They were also used

as a source of purple and black dyes. When Europeans were introduced to the plant, they brought

it home with them and cultivated it in Europe. During the 19th century, they were being grown

extensively in Russia for their oil. (https://www.hindawi.com/journals).

Due to huge and attractive flower heads, sunflowers are popular ornamental plants. Their

instantaneous growth and ease of cultivation making them a good choice for people who want to

do some gardening. Most garden stores carry an assortment of varieties, for people who want to

grow them in exotic colors like maroon and white.

The seeds have a high oil content ranging from 39-49% and are an important source of food

products. The oil is popular for cooking purposes and used in the production of margarine. It is

also high in unsaturated fats.

Heavy Metals

Heavy metals are natural components of Earths metal. They cannot be degraded or

destroyed. To a small extent, they enter our bodies through food, drinking water and air. As trace

elements, some heavy metals are essential to maintain the metabolism of the human body.

However, at higher concentration, they can lead to poisoning. Heavy metal poisoning could

result, for instance, come from drinking water contamination, high ambient air concentrations

near emission sources, or intake via the food chain.

Heavy metals are dangerous because they tend to bioaccumulate, which means there

wold be an increase in the concentration of a chemical in a biological organism over time,

compared to the chemicals concentration in the environment. Compounds accumulate in living

things any time they are taken up and stored faster than they are metabolized or excreted

(Lenntech 2002)

Lead

Lead is naturally-occurring element. It is harmful to humans when ingested or inhaled,

particularly to children under the age of six (United States Environmental Protection Agency).

For hundred of years, lead has been mined, smelted, refined, and used in products, including

gasoline and leaded pipes. Natural levels of lead in soil range between 50 ppm and 400 ppm

(United States Environmental Protection Agency). Lead a cumulative toxicant that affects mental
and physical development. Vital systems such as the circulatory and digestive systems are

affected. When exposed to lead, children, who are the most vulnerable, deals with problems

related to the nervous system (World Health Organization).

Lead Poisoning

Lead poisoning occurs when an organism absorb too much lead by breathing or

swallowing a substance with lead in it, such as paint, dust, water, or food. The activities of

humans vastly increased the concentration of lead on the surface of earth, putting our own

species at risk and poisoning plants and animals in the process. Contamination of aquatic

ecosystems occurs through extensive mining and industrial use of lead. Substantial amounts of

gasoline derived lead are deposited from the atmosphere into streams and other water bodies and

affect the animals and plants in these environment. Terrestrial plants and animals, some of which

share mans households, are also at great risk from contaminated soil. Lead can concentrate in

various kinds of plants, including single-celled algae in aquatic ecosystem, vegetables, grains

and fruit eaten by humans and domesticated animals. Shallow rooted plants, such as garden

plants and especially root crops such as carrots and radishes are more likely to become lead

contaminated (De pierie et.Al. 2003). The effects of lead on plants, especially at high

concentrations, are harmful . they include inhibition growth, interference with cell division and

with water absorption and balance, and reduction of photosynthesis, the vital process whereby

plants use the energy from sunlight to convert carbon dioxide and water into sugars, protein, fats

and other products. These translate into visible effects such as premature aging (browning of

leaves in optimal conditions), detachment of healthy leaves, noticeable, discoloration such as

back spots and darkening of veins on leaves and leaf damage. The by-products of photosynthesis
are useful in promoting growth and provide sustenance for the animal kingdom including

humans. Thus a reduction in the process of photosynthesis can cause harmful effects at multiple

levels in the food chain. Animals seem to be much more sensitive to lead than do the single

celled plants. Although lead is not an essential element for plants, it gets easily absorbed and

accumulated in different plant parts. Laboratory experiments of lead exposure carried out over a

28-day period in aquatic invertebrates (animals without backbones) including insects, snails and

amphipod crustaceans showed that in these animals that lead caused significant toxic effects.

Mammals are exposed to environmental lead and are affected by this neurotoxin in much the

same ways. A study in 1985 found neurotoxicity in monkeys exposed at levels that the Centres

for Disease Control is now calling dangerous to children. After exposure and initial high levels

of 25 mg/dL blood, stabilizing at a maximum of 13 mg/dL blood, the monkeys when tested

responded differently to controls to learning tasks, had impaired ability to learn a task involving

reversal of discrimination. Other researchers have observed similar decrements in learning ability

in laboratory rats exposed to lead. When lead is taken up in livestock, it accumulates mostly in

bone, which is a lead storage site and may contain 90% to 98% of total body burden. Bone-meal

is often added to other foods to increase phosphorus and calcium. Clinical lead-poisoning has

occurred in humans who have consumed such contaminated bone-meal as food supplement

(Demayo, Hodson et al. 1982). In humans, lead can damage almost every organ system

(WebMD). Elevated lead in the body can be detected by the presence of changes in blood cells

visible with a microscope and dense lines in the bones of children seen on X-ray, but the main

tool for diagnosis is measurement of the blood lead levels. When blood lead levels are recorded,

the results indicate how much lead is circulating within the blood stream, not the amount being

stored in the body. Exposure to lead leads to various negative effects on an organisms health
due to the fact that lead preferentially replaces other metals (e.g., zinc, calcium and iron) in

biochemical reactions. Lead interferes with the proteins that cause certain genes to turn on and

off by displacing other metals in the molecules, thus changing the shape of the protein molecule

such that it cant perform its function (Helemenstine, 2011). Effects of increased lead exposure

include damage to various systems of the body including the nervous and reproductive systems

and the kidneys, and it can cause high blood pressure and anaemia. It also interferes with the

metabolism of calcium and Vitamin D and at very high levels; it can cause convulsions, coma

and death in humans (World Health Organization, 2010). Other than humans, plants are also

affected by lead exposure (i.e. uptake). Although lead is not an essential element for plants, it

gets easily absorbed and accumulated in different plant parts. Excess lead in plants causes a

number of toxicity symptoms e.g. stunted growth and chlorosis.

Lead Contamination in Soil

For some state like Nebraska, Paint is a major contributor to soil lead contamination.

Remediation of exterior lead-based paint hazards is critical if further contamination is to be

avoided (Binder and Matte, 1992). The accumulation of lead in soil is primarily a function of the

rate of deposition from the atmosphere. The fate of lead in soil is affected by the specific or

exchange adsorption at mineral interfaces, the precipitation of sparingly soluble solid phases, and

the formation of relatively stable organo-metal complexes or chelates with the organic matter in

soil (EPA, 1986; NSF, 1977).

Evidence exists that atmospheric lead enters the soil as lead sulfate or is converted rapidly to lead

sulfate at the soil surface. Lead sulfate is relatively soluble, and thus could leach through the soil
if it were not transformed. In soils with pH of > or = 5 and with at least 5% organic matter,

atmospheric lead is retained in the upper 2-5 cm of undisturbed soil (EPA, 1986).

Lead may mobilize from soil when lead-bearing soil particles runoff to surface waters during

heavy rains. Lead may also mobilize from soil to atmosphere by downwind transport of smaller

lead- containing soil particles entrained in the prevailing wind (NSF, 1977). This latter process

may be important in contributing to the atmospheric burden of lead around some lead-smelting

and Superfund sites that contain elevated levels of lead in soil.

The downward movement of lead from soil by leaching is very slow under most natural

conditions (NSF, 1977). The conditions that induce leaching are the presence of lead in soil at

concentrations that either approach or exceed the sorption capacity of the soil, the presence in the

soil of materials that are capable of forming soluble chelates with lead, and a decrease in the pH

of the leaching solution (e.g., acid rain) (NSF, 1977). Partial favorable conditions for leaching

may be present in some soils near lead- smelting and NPL sites that contain elevated levels of

lead in soil.

Methodology

Materials

The Soil and the Sunflower seeds will be obtained from Bureau of Plant Industry. Soil will be

analyzed by it pH and Lead level

Biosorption Study

Sunflower seeds will be sown in tested soil. After germinating metal solutions of different

concentrations - lead 0, 100, 200, 300mg/kg of soil will be added to the plants. The plants will be
allowed to grow under suitable conditions like sunlight and moisture. The plants will be watered

twice a day with distilled water. After 20 days of incubation with metals, plants will be removed

and subjected to digestion.

Digestion of plants

The sunflower plant will be ashed using Hot air oven at 80c overnight. The ashed plant parts

will be put in separate test tubes. To these test tubes 5ml of Concentrated Nitric acid and 3ml of

Concentrated Perchloric acid will be put. These test tubes will be kept in Hot Air Oven for 5

minutes at 60c. After heating, the samples will be diluted using Diluted Nitric acid and will be

filtered to get clear solution. After filtering, the volume of the solution will be made up to 25ml

using Diluted Nitric acid tested for metals using AAS.

Statistical Treatment

T-test for two independent means will be used to test if sunflower can absorb lead metals

significantly .

Bibliography

[1] Angelova, V. et al. (2016) Potential of Sunflower (Helianthus annuus L.) for

Phytoremediation of Soils Contaminated with Heavy Metals. International Journal of Biological,

Biomolecular, Agricultural, Food and Biotechnological Engineering Vol:10, No:9

[2] Boominathan, R, et al. (2004) Production of Nickel Bio-Ore from Hyperaccumulator Plant

Biomass: Applications in Phytomining. Biotechnology and Bioengineering 86(3):243-50

[3] Fulekar, M.H. (2016) Phytoremediation of Heavy Metals by Helianthus annuus in Aquatic

and Soil environment. International Journal of Current Microbiology and Applied Sciences

ISSN: 2319-7706 Volume 5 Number 7 (2016) pp. 392-404

[4]Adesodun, J.K. et al. (2010) Phytoremediation Potentials of Sunflowers (Tithonia diversifolia

and Helianthus annuus) for Metals in Soils Contaminated with Zinc and Lead Nitrates. Water,

Air, and Soil Pollution March 2010, Volume 207, Issue 1-4, pp 195-201

[5] Alves, J.D. et al. (2016) Potential of sunflower, castor bean, common buckwheat and vetiver

as lead Phytoaccumulators.Revista Brasileira de Engenharia Agrcola e Ambiental ISSN 1807-

1929 v.20, n.3, p.243249

[6] Matthew, A.M. (2005) Phytoremediation of Heavy Metal Contaminated Soil Cochin

University of Science and Technology Cochin, Kerala, India

[7] Meers, E., A. Ruttens, M. Hopgood, E. Lesage and F.M.G. Tack. (2005) Potential of Brassica

rapa, Cannabis sativa, Helianthus annuus L., and Zea mays L., for phytoextraction of heavy

metals from calcareous dredged sediment derived soils. Chemosphere, 4: 561-572.

[8] Raskin, I., R.D. Smith and D.E. Salt. (1997). Phytoremediation of metals: Using plants to

remove pollutants from the environment. Curr. Opin. Biotechnol., 8: 221-226.

[9] Boonyapookana, B., Parkpian, P., Techapinyawat, S., Delaune, R.D., Jugsujinda, A. (2005).

Phytoaccumulation of lead by Sunflower (Helianthus annuus), Tobacco (Nicotiana tabacum) and

Vetiver (Vetiveria zizanioides). J. Environ. Sci. Health, 40: 117-137.

[10] Phytoremediation: An Environmentally Sound Technology for Pollution Prevention, Control

and Redmediation. Newsletter and Technical Publications Freshwater Management Series No. 2

from http://www.unep.or.jp/Ietc/Publications/Freshwater/FMS2/1.asp

[11] Klassen, S.P., McLean, J.E., Grossel, P.R., Sims, R.C. (2000). Fate and behavior of lead in

soils planted with metal resistant species (River birch and smallwing sedge), J. Environ. Quality,

29: 1826-1834.

[12] R. Ullah , J. Bakht, M. Shafi, M. Iqbal, A. Khan and M. Saeed (2011) Phytoaccumulation of

heavy metals by sunflower (Helianthus annuus L.) grown on contaminated soil. African Journal

of Biotechnology Vol. 10(75), pp. 17192-17198

[13] Karrari et al (2012) A systematic review on status of lead pollution and toxicity in Iran;
Guidance for preventive measures retrieved from
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3514537/