Comparative Study on the Lead Content of Moringa oleifera (Malunggay) from the Different

Geographical Locations in Luzon

OBJECTIVES

Generally, the study aims to compare the lead content of Moringa oleifera from the
following areas:
A. Metro manila- Valenzuela, Manila, Quezon
B. Suburban areas- Bulacan, Pampanga, Laguna

Specifically, the study aims to answer the following:

A. To determine the lead content of malunggay from the different areas using atomic
absorption spectra after extraction.
B. To compare the lead content of malunggay from the different areas using statistical
tools such as ANOVA.

BACKGROUND

Moringa oleifera is called Malunggay in Philippines and Moringa in english. The plant
grows wildly in hot tropical climate and is a wonderful herb known all over the world, but only
recently in the U.S. This plant is best known as an excellent source of nutrition and a natural
energy booster. Loaded with nutrients, vitamins and amino acids, it replenishes the human body
which is a remarkable discovery that can make a tremendous difference in the health and quality
of life. Mounting scientific evidence shows what has been known for thousands of years by
people in the tropical parts of the world. Yet this is not a sugar-based energy. It’s not something,
which makes the body hyper for some period of intense activity then leaves it drained. In fact,
Moringa is also relaxing where it helps reduce the blood pressure and assure a good night’s
sleep.

University laboratories around the world have studied Moringa’s ability to purify water,
attaching itself to harmful material and bacteria, and allowing them to be expelled as waste. It
produces long-lasting energy without hyperactivity. Other health benefits identified by people
who used Moringa stated the same pattern: immune system strengthened, skin condition restored,
blood pressure controlled, headaches and migraines handled, diabetes sugar level managed,
inflammations and arthritis pains reduced, tumors restricted and ulcers healed. Moringa is known
to be loaded with nutrients. Each ounce of Moringa contains seven times the Vitamin C found in
oranges, four times the Vitamin A of carrots, three times the iron of spinach, four times much
calcium as milk and three times the potassium of bananas. One of the best attributes of the
Moringa plant is that it is also quite tasty and a welcome addition to your kitchen. The leaves,
pods and flowers of this versatile tree are all edible, each with its own flavor. They can be
served fresh with meals, or be reduced to powder and used as a food supplement.
 With so much nutritional aid Moringa can give to our life as a whole, we would like to
make a comparative study on lead content of Moringa from different geographical areas. This
study will make us aware of the lead content of Moringa. We aim to determine and compare the
lead content of Moringa from the following areas:

a. Metro Manila – Valenzuela, Quezon City and Manila

b. Provinces - Laguna, Pampanga, Bulacan

The most serious source of exposure to soil lead is through direct ingestion (eating) of
contaminated soil or dust. In general, plants do not absorb or accumulate lead. However, in soils
testing high in lead, it is possible for some lead to be taken up. Studies have shown that lead does
not readily accumulate in the fruiting parts of vegetable and fruit crops (e.g., corn, beans, squash,
tomatoes, strawberries, apples). Higher concentrations are more likely to be found in leafy
vegetables (e.g., lettuce) and on the surface of root crops (e.g., carrots).

Lead is the heaviest of the non-radioactive metals (atomic number 82; atomic weight
207) that naturally occur in substantial quantities in the earth's surface. Lead is present in all
soils, rivers, lakes, and seawater. Despite its weight, lead is also in the air, a component of dust
and of sea spray. Lead is present in the proportion of 16 parts per million (ppm) in igneous rocks,
the most common ancient rock on the surface, and at an average of about 10 ppm in common
soils that are far from sites of contamination; natural soils usually have less than 50 ppm of lead
but are never lead free.

The main health problems associated with chronic high levels of lead in the blood are
neurological impairment in children (including possible permanent damage if exposed to high
levels at an early age) and hypertension in adults. Pregnant women must be especially cautious
about lead exposure, both because of increased risk of spontaneous abortion and potential
damage to the fetus.

Lead first enters the food supply in significant quantities by contamination of the soil and
by lead dust accumulating on the plants. The lead is taken up by plants through roots and leaves.
The plant materials are eaten by humans, and animals that are eaten by humans gain lead from
the plants they eat. Although proponents of vegetarian diets claim that meat is particularly
contaminated by heavy metals, plant foods tend to have higher lead levels than animal foods
because they can absorb and retain more lead than most animals. Animals have a low absorption
rate of ingested lead and have physiological mechanisms to discharge the ingested lead; much is
sequestered into the bones, which are usually not consumed by humans. By contrast plants are
efficient at absorbing soil lead and retain the lead that has been taken up. About 7% of the lead in
soil is taken up by plants (excessive lead will kill off the plants).

Lead from the atmosphere that lands on soils has low mobility and tends to stay in the top
inch of soil. Therefore, shallow-rooted plants, such as grasses and common vegetables, are
particularly vulnerable to picking up lead contamination that originated in the atmosphere.
 The human body eliminates lead via many mechanisms. Lead exits the body via: feces
(mainly as the result of dietary lead not being absorbed, but also discharged via the gallbladder
from breakdown of hemoglobin that binds lead); through sweat; excretion into the skin, hair, and
nails; and through the urine and the breath. Except at times of unusually high lead exposure, the
total lead taken into the body each day is eliminated each day. A certain relatively constant level
of lead remains in circulation in the blood; small amounts are also present in all the soft tissues
and larger amounts are present in the bones; the levels depend on the average daily exposure to
lead. The foods could become contaminated in these environments with impure water used in
preparation and by dust and dirt that contained high levels of lead.

REVIEW OF LITERATURE

The most serious source of exposure to soil lead is through direct ingestion (eating) of
contaminated soil or dust. In general, plants do not absorb or accumulate lead. However, in soils
testing high in lead, it is possible for some lead to be taken up. Studies have shown that lead does
not readily accumulate in the fruiting parts of vegetable and fruit crops (e.g., corn, beans, squash,
tomatoes, strawberries, and apples). Higher concentrations are more likely to be found in leafy
vegetables (e.g., lettuce) and on the surface of root crops (e.g., carrots).

Since plants do not take up large quantities of soil lead, the lead levels in soil considered
safe for plants will be much higher than soil lead levels where eating of soil is a concern (pica).
Generally, it has been considered safe to use garden produce grown in soils with total lead levels
less than 500 ppm. The risk of lead poisoning through the food chain increases as the soil lead
level rises above this concentration. Even at soil levels above 500 ppm, most of the risk is from
lead contaminated soil or dust deposits on the plants rather than from uptake of lead by the plant.

Because of the possibility of bare soil exposure to children through hand to mouth
activity, soils with lead levels exceeding 100 ppm should not be used for gardening. If soil
exposure to children is not a concern, then plants can be safely eaten from soils with soil lead
levels up to 300 ppm for leafy and root type vegetables, and 500 ppm for fruits and vegetables
where the fruiting parts are eaten.

Lead, a highly hazardous yet poorly monitored substance, have a long half life. Its
movement in the atmosphere through vehicular emissions and its deposition in different
environmental elements will be a global burden for a long time (Landstown 1986). Leaded
gasoline was phasedout in Metro Manila in sometime April of 2000 with total phase out in the
whole of the archipelago in December of the same year. Such move was partly due to the
implementation of clean air Act of 1999 as well as the environmental concern of previous
president Fidel V. Ramos (The WorldBank Group 2002). Though gasoline is no longer leaded in
the Philippines, still children have considerable blood lead levels and such is considered a public
health concern (Subida 2003). Lead abounds in different areas and depot. Foreign studies
showed that lead may be accumulated in soil.

A study in the United Kingdom revealed that lead levels in soil increases with increasing
number of vehicles passing that area. The same has been observed with plants in Berlin (Klumpp
2003). In Hungary, airborne lead concentrations along the main road in Gyor were found to be 3-
6 times higher than the hygiene limit of 3 ug/m3.. The differences in the children’s lead exposure
in the different areas of the town were reflected in the differences in lead content of the soil in
the neighborhoods of the children's schools (WHO 1972). Lead thus may be deposited in plants
and soil with resulting adverse effects to man.

Significant increases in the Pb content of cultivated soils have been observed near
industrial areas. Pb tends to accumulate in the surface ground layer and its concentration
decreases with soil depth (de Abreu et al., 1998). It is easily taken up by plants from the soil and
is accum ulated in different organs. Pb is considered a general protoplasmic poison, which is
cumulative, slow acting and subtle. Soils contaminated with Pb causes sharp decreases in crop
productivity thereby posing a serious problem for agriculture (Johnson and Eaton, 1980). The
present review focuses on the sources of Pb; its uptake and transport within plant; physiological,
biochemical and ultrastructural changes due to Pb toxicity; Pb tolerance in plants as well as
possible remediation measures for Pb contaminated soils.

Pb is a major pollutant in both terrestrial and aquatic ecosystems. Besides natural

weathering processes the main sources of Pb pollution are exhaust fumes of automobiles,
chimneys of factories using Pb, effluents from the storage battery, industry, mining and smelting
of Pb ores, metal plating and finishing operations, fertilizers, pesticides and additives in pigments
and gasoline (Eick et al., 1999). Sewage sludge containing large quantities of Pb and other
metals is regularly discharged on to field and garden soils due to increasing trends in
urbanization (Paivoke, 2002). The accumulated Pb on the street and highways is transported to
surface streams by rain water and consequently pollutes others surface waterways and soil
(Laxen and Harrison, 1977). Compounds of Pb used as agricultural chemicals such as Pb
arsenate, which is used as a pesticide, contaminate agricultural soils.

Pb is available to plants from soil and aerosol sources. Pb uptake studies in plants have
demonstrated that roots have an ability to take up significant quantities of Pb whilst
simultaneously greatly restricting its translocation to above ground parts (Lane and Martin,
1977). This notion was overturned by Miller and Koeppe (1971) who demonstrated that Zea
mays L. plants could translocate and accumulate significant quantities of Pb in the leaves in a
concentration dependent manner. The extent to which Pb enters plants via the leaves depends on
the ability of leaves to absorb Pb from aerial sources, which in turn depends on the specific leaf
morphology. Downy leaves absorb heavy metals from the atmosphere (Godzik, 1993). However
it is agreed that the bulk of the Pb taken up by plants remains in the roots (Kumar et al., 1995).
Pb accumulates in the surface layers of soils and therefore it is difficult to reliably measure the
portion of soil Pb directly available to plants. Its availability depends highly on soil conditions.
Pb binds to organic material in the soil. Soil particle size and cation exchange capacity as well as
plant factors such as root surface area, root exudates, mycorrhization and rate of transpiration
affect the availability and uptake of Pb (Davies, 1995). The absorption of Pb (as Pb) in soil
follows the Langmuir relation and increases with increasing pH between 3.0 to 8.5 (Lee et al.,
1998). However Blaylock and coworkers (1997) reported that in soil with a pH between 5.5 and
7.5 Pb solubility is controlled by phosphate or carbonate precipitates and very little Pb is
available to plants even if they have the genetic capacity to accumulate it. Pb in soil is classified
as a weak Lewis acid, which implies a strong covalent character to many of the ionic bonds it
forms in soils and plants.
 Pb present in the soil is nearly always tightly bound to organic or colloidal material or in
a precipitated form, all of which serve to reduce the uptake of Pb by plant roots.

There is more concern about lead contamination from external lead on unwashed produce
than from actual uptake by the plant itself if the location is on the highways or areas where
pollution is predominantly present.

The "Moringa" tree is grown mainly in semi-arid, tropical, and subtropical areas,
corresponding in the United States to USDA hardiness zones 9 and 10. While it grows best in dry
sandy soil, it tolerates poor soil, including coastal areas. It is a fast-growing, drought-resistant
tree that is native to the southern foothills of the Himalayas in northwestern India. Reports that it
grows wild in the Middle East or Africa are completely unsubstantiated. Today it is widely
cultivated in Africa, Central and South America, Sri Lanka, India, Mexico, Malaysia, and the
Philippines. It is considered one of the world’s most useful trees, as almost every part of the
Moringa tree can be used for food or has some other beneficial property. In the tropics, it is used
as forage for livestock, and in many countries, Moringa is used as a micronutrient powder to treat
diseases.

The immature green pods called “drumsticks” are probably the most valued and widely
used part of the tree. They are commonly consumed in India and are generally prepared in a
similar fashion to green beans and have a slight asparagus taste. The seeds are sometimes
removed from more mature pods and eaten like peas or roasted like nuts. The flowers are edible
when cooked, and are said to taste like mushrooms. The roots are shredded and used as a
condiment in the same way as horseradish; however, it contains the alkaloid spirochin, a
potentially fatal nerve-paralyzing agent, so such practices should be strongly discouraged.

The leaves are highly nutritious, being a significant source of beta-carotene, Vitamin C,
protein, iron, and potassium. The leaves are cooked and used like spinach. In addition to being
used fresh as a substitute for spinach, its leaves are commonly dried and crushed into a powder,
and used in soups and sauces. Murungakai, as it is locally known in Tamil Nadu and Kerala, is
used in Siddha medicine. Its leaves are full of medicinal properties. The tree is a good source for
calcium and phosphorus. In Siddha medicine, the drumstick seeds are used as a sexual virility
drug for treating erectile dysfunction in men and also in women for prolonging sexual activity.

The Moringa seeds yield 38–40% edible oil (called ben oil from the high concentration of
behenic acid contained in the oil). The refined oil is clear, odorless, and resists rancidity at least
as well as any other botanical oil. The seed cake remaining after oil extraction may be used as a
fertilizer or as a flocculent to purify water.

The bark, sap, roots, leaves, seeds, oil, and flowers are used in traditional medicine in
several countries. In Jamaica, the sap is used for a blue dye. The flowers are also cooked and
relished as a delicacy in West Bengal and Bangladesh, especially during early spring. There it is
called sojne ful and is usually cooked with green peas and potato.
 Moringa trees have been used to combat malnutrition, especially among infants and
nursing mothers. Three non-governmental organizations in particular — Trees for Life, Church
World Service, and Educational Concerns for Hunger Organization — have advocated Moringa
as “natural nutrition for the tropics.” Leaves can be eaten fresh, cooked, or stored as dried
powder for many months without refrigeration, and reportedly without loss of nutritional value.
Moringa is especially promising as a food source in the tropics because the tree is in full leaf at
the end of the dry season when other foods are typically scarce. (Jed W. Fahey, 2005)

A large number of reports on the nutritional qualities of Moringa now exist in both the
scientific and the popular literature. It is commonly said that Moringa leaves contain more
Vitamin A than carrots, more calcium than milk, more iron than spinach, more Vitamin C than
oranges, and more potassium than bananas,” and that the protein quality of Moringa leaves rivals
that of milk and eggs. However, the leaves and stem of M. oleifera are known to have large
amounts of their calcium bound in calcium oxalate crystals, (Olson, M. E., and S. Carlquist.
2001. Stem and root anatomical correlations with life form diversity, ecology, and systematics in
Moringa (Moringaceae), Botanical Journal of the Linnean Society 135(4): 315-348), which is not
a form of calcium available to the body. Whether the claim of "more calcium than milk" includes
this non-bioavailable calcium needs to be addressed. The oral histories recorded by Lowell
Fuglie in Senegal and throughout West Africa, who reports countless instances of lifesaving
nutritional rescue that are attributed to Moringa (Fuglie, L.J., 1999, 2000). In fact, the nutritional
properties of Moringa are now so well-known that there seems to be little doubt of the substantial
health benefit to be realized by consumption of Moringa leaf powder in situations where
starvation is imminent. Nonetheless, the outcomes of well-controlled and well-documented
clinical studies would still be clearly of great value. (Jed W. Fahey, 2005) In many cultures
throughout the tropics, differentiation between food and medicinal uses of plants (e.g. bark, fruit,
leaves, nuts, seeds, tubers, roots, and flowers), is very difficult because plant uses span both
categories, and this is deeply ingrained in the traditions and the fabric of the community (Lockett
et al., 2000). ”

MATERIALS AND METHODS

The researchers will be using the random sampling method in order to collect the leaves
of Malunggay plant (Moringa oleifera). Six different places will be used to conduct the said
sampling: 50g of Malunggay leaves each in three different places in Metro Manila specifically
Manila, Quezon City and Valenzuela City and three different places in provinces such as
Bulacan, Laguna, and Pampanga. After the collection was performed, the leaves will be then
subjected to undergo into extraction.
Hydro Distillation extraction method will be performed to get the liquid component of
the plant. In this method, the plant material is charged in the still and the water is added to
immerse the charge, leaving sufficient vapor space. The quantity of water should be adequate for
the plant material to move freely in boiling water and avoiding localized overheating. The water
is heated under direct fire or steam jacket or in some cases a closed steam coil. Water will be
added during the distillation process to prevent dry material from being exposed to direct
heating. The steam vapor produced is condensed and oil is separated from the water as in steam
distillation and each extracted liquid samples will be placed in an Erlenmeyer’s flask.
 After the extraction method, it will be then subjected to Atomic Absorption Spectroscopy
(AAS) to determine the concentration of a particular metal element within the said research
sample: the presence of Lead (Pb) in the extracts of Malunggay leaves. This AAS can be also
used to analyze the concentration over 62 different metals in a solution.
After Atomic Absorption Spectroscopy test is done, the different samples will be
analyzed using the ANOVA (Analysis of Variance) test and will be then compared in a tabulated
form.

REFERENCES
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http://pharmacybd.com/download/assignments/Atomic Absorption Spectroscopy.doc
2. Pati, Kumar. Malunggay. Best Nutrition.
http://www.malunggay.com/description.htm
3. Rajangam J., et al. Status of Production and Utilisation of Moringa in Southern India. In
"Development potential for Moringa products", October 29th - November 2nd, 2001, Dar es
Salaam, Tanzania.
4. Samara Company. 2008. AromaticPlantExtractionMethods. Samara Botany: Nature Intelligence.
http://www.wingedseed.com/aromatherapy_information/AromaticPlantExtractionMethods.htm
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http://www.itmonline.org/arts/lead.htm