Dulaw as Dye

CHAPTER I
INTRODUCTION

Background of the Study
Round Turmeric (Cucurma Domestica Valet) is commonly known in our community by the name “dulaw”. The plant is native to Southern Asia. The parts of this plant used medicinally are the rhizome and the root. Turmeric is a close relative to Ginger Root, and is used as a cooking spice in India & other Asian countries. It has been used in traditional medicine for the treatment of jaundice and other liver ailments, ulcers, parasitic infections, various skin diseases, sprains, strains, bruises, inflammation of the joints, cold and flu symptoms, preserving food, and promoting digestion. Native peoples of the Pacific sprinkled the dust on their shoulders during ceremonial dances, and used it for numerous medical problems ranging from constipation to skin diseases.
A perennial plant with roots or tubers oblong, palmate, and deep orange inside; root-leaves about 2 feet long, lanceolate, long, petioled, tapering at each end, smooth, of a uniform green; petioles sheathing spike, erect, central, oblong, green; flowers dull yellow, three or five together surrounded by bracteolae. It is propagated by cuttings from the root, which when dry is in curved cylindrical or oblong tubers 2 or 3 inches in length, and an inch in diameter, pointed or tapering at one end, yellowish externally, with transverse, parallel rings internally deep orange or reddish brown, marked with shining points, dense, solid, short, granular fracture, forming a lemon yellow powder. It has a peculiar fragrant odor and a bitterish, slightly acrid taste, like ginger, exciting warmth in the mouth and colouring the saliva yellow. It yields its properties to


water or alcohol. As a culinary herb, the stimulating taste of Turmeric adds zip to foods. As a
healthful herbal tonic, it is said to provide warming, soothing benefits to the joints, liver, and stomach. Women also appreciate its calming benefits during periods of monthly discomfort. (http://www.mdidea.com/products/new/new088.html#01)

From the classifications above, natural dye will be the end product of this research, because the source of dye is a rhizome from Round Turmeric .And so, we got the idea of making a study out of a dye from Round Turmeric when accidentally while treating various infection using the rhizome wherein it stained our white shirts.

Statement of the Problem
Dye is used to color objects or textiles and as of now dyes are getting expensive.
This study was conducted to extract dye from rhizomes of Round Turmeric.
1. Is there any significant difference between the color produced from Round Turmeric dye and mangosteen dye?
2. Is there any significant difference between the dye quality of Round Turmeric and Mangosteen?
3. What is the color of the extracted dye?
4. What type of cloth will the dye retain its color more, polyester, cotton, gina earth, or in satin after bleaching, rubbing with running water, and sun exposure?

Hypothesis
1.There will be a difference between the dye of Round Turmeric and Mangosteen dye.
2. There will be a difference between the dye quality of Round Turmeric and Mangosteen.
3. The color of the extracted dye is dark yellow.
4. The dye retains more on the cotton and polyester clothes after bleaching, rubbing with running water and sun exposure.

Significance of the study
Today, prices of foods and other accessories keep on increasing. One of these
things is dye. The researchers attempt to find some alternative sources for cheaper cost.
This study aims to prove if round turmeric can change the color of the clothes. If this research will succeed in producing high quality natural dyes, then this would be a great help for low-income families.

Scope and Delimitations
This research dealt with the extraction of Round Turmeric rhizomes and comparing it to Mangosteen dye. It involves with a mordant bath, money, time, and especially round turmeric was the components to succeed in the round turmeric dye making which will be conducted at ESEP Building at the laboratory of Iligan City National High School (ICNHS) S.Y. 2007-2008.
This study will only limit on producing dye out of rhizomes from round turmeric (dulaw), comparing it to mangosteen dye and in testing the products quality with people who are high in color sensitivity specifically females because color blindness is typically a genetic condition, and it is much more common in men than in women. Approximately one in 12 men has at least some color perception problems. Less common, acquired deficiencies stem from injury, disease, or the aging process. Also, although not called "color blindness," when people age; their corneas typically turn yellowish, severely hampering their ability to see violet and blue colors. (http://webexhibits.org/causesofcolors/2C.html)

The researchers also limit their study by using mangosteen dye not a commercial dye as their control variable since mangosteen’s chemical properties, physical properties and its composition are already known to them.

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CHAPTER II
Review of Related Literature and Studies

Review of Related Literature

Round Turmeric
Round Turmeric (Cucurma Domestica Valet) is commonly known in our community as “Dulaw”. It is herbaceous perennial plant belonging to the natural order Zingibreae. It is native of Southern Asia, and being cultivated on a large scale. Round Turmeric, representing the main rhizomes is ovate or pear shaped, about the size of a pigeon’s egg, and marked externally with the numerous annular wrinkles. Sometimes it comes cut into transverse segments. Turmeric is used principally as a constituent of curry powders and other condiments and is already a part of the cultures of most Muslim in the Philippines. The contents of many of the cells becoming swollen to form pasty masses which are colored yellow by the curcumin present in the cells. It has been employed in the treatment of chronic cholecystitis. Also, it grows in all kinds of soil and is suitable to our environment. Round Turmeric has been used from remote areas as a condiment and to a more limited extent as a medicine. It is also used for bruises.
Used in the recommended amounts, turmeric is generally safe. It has been used in large quantities as a condiment with no adverse reactions. While some herbal books recommend not taking high amounts of turmeric during pregnancy as it may cause uterine contractions and people with gallstones or obstruction of bile passages should consult their healthcare practitioner before using turmeric. Round Turmeric Dye which is a natural dye has no side effects on skin.
(http://www.evitamins.com/healthnotes.asp?ContentID=2175005)


Dye
Dyes give color to the textile, fabrics or clothes and to other materials including some foods. Dyes can be natural or synthetic. Natural Dye can be obtained from vegetable, mineral resources, and plants. A dye can generally be described as a colored substance that has an affinity to the substrate to which it is being applied. The dye is generally applied in an aqueous solution, and may require a mordant to improve the fastness of the dye on the fiber. Both dyes and pigments appear to be colored because they absorb some wavelengths of light preferentially. In contrast with a dye, pigment is generally is insoluble, and has no affinity for the substrate. Some dyes can be precipitated with an inert salt to produce a lake pigment.
“Dye is a colored substance used to impart more or less permanent color to other substances”. Most of the dyes are used in clothing and textiles where it passes in a long process. Humans have applied color to clothing for nearly all of recorded history including Egyptians, Indians, Chinese, and other ancient people. Usually in these days dye making or dyeing needs machines to come up with the dye but here, in this research only the simpliest and the easiest process is used. Dyes are obtained from wood, barks, flowers, leaves, fruits and seeds of various plants. The country is compelled to import for about $20 million worth of dyes, tannins, and printing pigments annually due to lack of local manufacturers of either synthetic or natural dyestuffs.
According to Groiler Childrens Encyclopedia (1994, p.215) most dyes are now made from chemicals. To dye an object, dip it in water containing dissolved dye. If the dye is fast, the object will keep its dyed color no matter how often you wash it.” Dyes are generally fast, that's why they retain their color in the fiber throughout the dyeing process. There are types of dyes.

First and the most common is the natural dye which is made from plants and animals. Second is the mineral dye made from materials dug from the earth. A synthetic dye which is composed of man made chemical substances is the last and least used. Synthetic Dyes are toxic it can create allergic reaction to skin and contaminate the environment. Because of the disadvantages of synthetic dye, natural dyestuff has revived in Europe, Japan, and U.S.
(http://www.hort.cornell.edu/plantsandtextiles/pdfs/indigo/introduction.pdf)

Curcumin
The curcumins are polyphenols and are responsible for the yellow color of turmeric. Curcumin can exist in at least two tautomeric forms, keto and enol. Since curcumin is brightly colored, it may be used as a food coloring. As a food additive, its E number is E100.
The active constituent known as curcumin has been shown to have a wide range of therapeutic actions. First, it protects against free radical damage because it is a strong antioxidant. Second, it reduces inflammation by lowering histamine levels and possibly by increasing production of natural cortisone by the adrenal glands. Third, it protects the liver from a number of toxic compounds. Fourth, it has been shown to reduce platelets from clumping together, which in turn improves circulation and may help protect against atherosclerosis. (http://www.evitamins.com/healthnotes.asp?ContentID=2175005)
Curcumin, the major constituent of turmeric is a known antioxidant. We have examined the oxidative folding of the model four-disulfide-bond-containing protein bovine pancreatic ribonuclease A (RNase A) in its presence; results indicate that RNase A regeneration rate increases in a curcumin-dependent manner. Examination of the native tendency of the fully-reduced polypeptide and the stability of key folding intermediates suggests that the increased oxidative folding rate can be attributed to native-like elements induced within the fully-reduced polypeptide and the stabilization of native-like species by this non-redox-active natural product. Our results provide a template for the design of curcuminoid-based synthetic small-molecule fold catalysts that accelerate the folding of ER-processed proteins; this assumes significance given that nitrosative stress and dysfunction of the ER-resident oxidoreductase protein disulfide isomerise due to S-nitrosylation are factors associated with the pathogenesis of Alzheimer's and Parkinson's diseases. (http://www.ncbi.nlm.nih.gov/sites/entrez?cmd=Retrieve&db=PubMed&dopt=AbstractPlus&list_uids=17959149)
Curcumin has been shown to exhibit anti-inflammatory, antimutagenic, and anticarcinogenic activities. However, the modulatory effect of curcumin on the functional activation of primary microglial cells, brain mononuclear phagocytes causing the neuronal damage, largely remains unknown. The current study examined whether curcumin influenced NO production in rat primary microglia and investigated its underlying signaling pathways. Curcumin decreased NO production in LPS-stimulated microglial cells in a dose-dependent manner, with an IC50 value of 3.7 μM. It also suppressed both mRNA and protein levels of inducible nitric oxide synthase (iNOS), indicating that this drug may affect iNOS gene expression process. Indeed, curcumin altered biochemical patterns induced by LPS such as phosphorylation of all mitogen-activated protein kinases (MAPKs), and DNA binding activities of nuclear factor-κB (NF-κB) and activator protein (AP)-1, assessed by reporter gene assay. By analysis of inhibitory features of specific MAPK inhibitors, a series of signaling cascades including c-Jun N-terminal kinase (JNK), p38 and NF-κB was found to play a critical role in curcumin-mediated NO inhibition in microglial cells. The current results suggest that curcumin
is a promising agent for the prevention and treatment of both NO and microglial cell-mediated neurodegenerative disorders. (http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6T99-4KR3JRN-1&_user=10&_coverDate=10%2F19%2F2006&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000050221&_version=1&_urlVersion=0&_userid=10&md5=4cd385a5c1de1dcfed135c0fd7b8055b)
Mordant
A mordant is a substance used to set dyes on fabrics by forming an insoluble compound with the dye. It may be used for dyeing fabrics, or for intensifying stains in cell or tissue preparations. A mordant is either inherently colloidal or produces colloids and can be either acidic or alkaline.
Mordants include tannic acid, alum, chrome alum, sodium chloride, and certain salts of aluminum, chromium, copper, iron, iodine, potassium, sodium, and tin. Iodine is used as a mordant to set the first dye in gram stains. Phosphomolybdic acid is also used as a mordant to set light green when staining with Masson's trichrome stain. Mordants are two part "developed" dyes used to dye cotton with dark colors. The initial bath imparts a yellow or pale chartreuse color. This is oxidized in place to produce the brownish we are familiar with in rusts.It is important to note that many mordants, particularly those in the hard metal category, can be hazardous to health and extreme care must be taken in using them. (http://www.evitamins.com/healthnotes.asp?ContentID=2175005)

Gallic Acid
Gallic acid is created by the hydrolysis of gallotannic acid. Gallotannic acid consists of a glucose molecule with 5 gallic acid groups or di-gallic acid groups in any combination. Galls contain a high concentration of gallotannic acid but less of gallic acid. To increase the gallic acid concentration, the gallotannic acid can be hydrolyzed during preparation.of the ink. This can be achieved best by boiling the gallotannic acid in an acidic environment or by letting the gallotannic acid solution ferment. Enzymes released by the mold react with the glucose and hydrolyze the gallic acid in the process.

The iron(II) ions of the vitriol react with both gallic acid and gallotannic acid. The precise chemical structure of the iron gall ink pigment has recently been investigated by two researchers (C.H. Wunderlich and C. Krekel) and each has developed different theories on the structure of the colored compound.
Wunderlich was able to form black crystals using Fe(III)Cl3 and gallic acid in a gel of sodium silicate. He suggests that the iron reacts with the three hydroxyl groups of gallic acid and with the carboxyl group, creating a three dimensional structure. The color formation in this ferric tannate complex is due to shifting electron pairs in the benzene ring and oxygen/iron bonds. Using Mossbauer and XANES spectroscopy he was able to determine an oxidation state (Fe 3+). Wunderlich created these molecules using Fe(III) ions. However, when iron gall ink is produced using iron sulfate, the ferrous tannate complex is formed with Fe(II).
Krekel has researched pigment formation using different metals under varying circumstances. He was able to create a black pigment which he examined using different analysis techniques (mass spectrometry, infrared spectrometry and Mossbauer spectrometry). His study suggests that the black pigment, instead of an iron gallic acid complex, is an iron pyrogallol complex. (http://www.knaw.nl/ecpa/ink/ink_chemical.html)
Gallic acid can bound with a metal such as copper as shown by this structure. (http://pubs.acs.org/isubscribe/journals/joceah/65/i21/figures/jo000335zh00005.html)

This structure shows that curcumin (diferuloylmethane) belonging to the same organic group as the gallic acid can also bind with metal such as Fe+2. (http://en.wikipedia.org/wiki/Curcumin)

Review of Related Studies
Indigo Dyeing
Indigo works by a chemical reaction called oxidation-reduction. Indigo does not dissolve in water. It must be reduced — i.e. the oxygen must be removed— in the presence of alkali by a reducing agent such as thiourea dioxide (thiox), sodium hydrosulfite, Zinc, or bacteria. Upon reduction, indigo becomes colorless and water soluble. In this state, indigo has a high affinity for cellulosic fibers and enters the open spaces of the fiber. The dyed fibers are then exposed to air, which oxidizes the dye molecule back to its insoluble form. The insoluble dye particles are trapped inside the fiber, coloring them permanently blue. Unlike most dyes, indigo forms a mechanical, not chemical, bond.
Indigo is a vat dye, so named because the traditional processing of indigo included fermenting the leaves in a vat (vessel). The fermentation process reduces indicant to its colorless, soluble form that fabric can absorb. To prevent premature oxidation, dyeing must take place in the absence of air. This fact dictates the technology of indigo dyeing. If one brushes the dye solution onto fabric, the brush might turn blue but not the fabric as the dye would become insoluble between the dye vat and the fabric. Traditional patterning of indigo-dyed fabrics usually depends on 1) fabric structure — mixing indigo yarns with other yarns, 2) physical resists that prevent dye penetration, 3) chemical resists that prevent dye oxidation, or 4) removal (discharge) of color after dyeing. A more recent option is the use of Inkodye, a soluble vat dye that has been preprocessed into the reduced form for direct application. (http://www.hort.cornell.edu/plantsandtextiles/pdfs/indigo/introduction.pdf)
“Tibor (1996) cited that a good dye must posses qualities such as colorfastness, long storage life and color intensity. He also cited that our country is blessed with vast natural resources, which can be utilized for sources of natural dyes.”
“Castillo (2002) also cited that when something is dyed, chemical or physical forces hold the color fast to the object. If these forces are not holding the color fast, the material is stained, not dyed. The chemical and physical forces are of different kinds and many dyes will stick firmly
to some materials but wash out of others easily”. Example, a blue colored dye holds firmly in cotton and red dye holds firmly on a silk fabric; however, the blue one fades faster on silk while the red one fades faster on cotton.”
“Lomansoc (2004) cited that in 1956, the introduction of reactive dyes for cotton, silk and wool revolutionized dyeing methods. While other dyestuffs form only loose bonds with fibers, reactive dyes form stable chemical links with textile materials to produce colored fabrics with excellent overall fastness.”
“Batu (2004) cited that in 1855, William Perkin, an 18-year-old chemistry student discovered a purple dye accidentally. The dye was not particularly fast but it became popular. Perkin called it mauve from the French word mallow. One of the most famous historical dye was Tyrian purple; a dye produced from shells of the marine mollusk Murex brandaris . This dye was expensive and in ancient Rome, it was reserved for the ruling class (hence the term “royal purple”).
“Gaid (2003) cited that extraction is the process used in organic chemistry to separate mixtures. Solvents are used in which the component to be extracted is highly soluble. After treatment of the mixture with the solvent which now contains the extracted material, the desired substance may be isolated by removal of the solvent through distillation or evaporation.”
Daro (2004) cited that the plant and animal fibers we use to make our textiles have little natural color. They are mostly off-white or brownish-yellow. We therefore have to add color to them in a form of a dye.
A dye must be dissolved before it can work. When textiles are placed into a dye bath (dye solution), the fibers absorb the molecules of the dye. These molecules give the fibers the desired colors.
A colorfast fabric does not change color under normal use. Such substance like chlorine bleach and perspiration may also affect the colors of fabrics.”
Gaid's research is all about extraction of oils or essential oil from oregano leaves. We cited a part in which he discusses the extraction process which improved our information about the extraction process.
Daro's research is also about dyes and dyeing. She discusses in the citation the fibers and textiles that is linked to dyeing. She also talked about the factor that affects the dye color in the textile. Her citation is also like Gaid's, the process of dye solution.
Tibor's research is about dyeing that's why we cited his citation. We all know that each organism is unique and because of that research's also has its own originality. Castillo's research is all about comparison of two kinds of dyes in one species or source called gemilina plant. Our research is about extraction of dye or finding a good source of dye from a rhizome. His /Her research is about comparing or comparison while ours is about extraction or production.
The research title of Lomansoc's research is “production of dye from the Sap of an avocado seed” while ours is “Extraction of Dye from Rhizomes”. In that case, production and extraction have almost the same meaning so we may expect that my research and
Lomonsoc's research has the same objectives. There may be difference but their objectives are similar.


Conceptual Framework

Null Hypothesis
1. There is no significant difference between the degree of yellow color from Round Turmeric and Mangosteen.
2. There is no significant difference between the cloth dyed with Round Turmeric and Mangosteen when bleached with detergent soap.
3. There is no significant difference between the cloth dyed with Round Turmeric and Mangosteen when ran with water.
4. There is no significant difference between the cloth dyed with Round Turmeric and Mangosteen when exposed to sunlight.
5. The color of the extracted dye is not dark yellow.
6. The dye did not retain more on the cotton and polyester clothes after bleaching, rubbing with running water and sun exposure.

Definition of Terms
Cotton - Thread or cloth manufactured from the fiber of these plants. (Cotton plant)
Discoloration - a change in the original or desired color of something that gives it an
unpleasant, faded, darkened, or dirty appearance.
Dye - is colored substance used to impart color textiles.
Dyeing - the use of the dye to change color of something permanently.
Fiber - fabrics thread
Linen - Cloth woven from the thread made from the fibers of the flax plant.
Mordant - Serving to fix colors in dyeing.
Quality test- used to measure the degree of change in color from white to yellow.

Satin - A smooth fabric, as of silk or rayon, woven with a glossy face and a dull back.


CHAPTER III
METHODOLOGY

Materials:
Round Turmeric Forceps
Clean Water Camera
Detergent Soap Beaker
Mortar and Pestle Dropper
Knife Pyrex
Clean Containers Gloves
Mordant (FeSO4) Mordant Cloths (cotton, gina earth, satin, polyester)

Gathering of Materials:
The researchers collected and gathered all needed materials and apparatuses for such experimentation. The materials were then washed thoroughly and were made sure that they were free from dirt. The quality of the materials that were used was secured because there may be damages or leaks that could greatly affect the experiment. In dyeing, four kinds of cloths will be used: cotton, gina earth, satin and polyester. Each of the fabrics should be 3 by 4 inches wide. And there should be 2 pieces of cloth of each kind.

Extraction of Dye from Round Turmeric
1. Gather as many “dulaw” as needed to produce at least 100ml. extract. In extracting, use clean rubber or gloves to avoid such dirt that may affect the quality of the dye.
2. Peel the “dulaw” and then place it into mortar. The peeled “dulaw” were then pounded using the pestle for about 15 minutes or until there is a liquid matter.
3. Place the liquid matter and the pounded “dulaw” on the pyrex with clean water filling the container half full.
4. Boil the liquid and residue on a stove for about 15 minutes.
5. Filter the boiled residue leaving the liquid one in a clean container.
6. Cool the dye for 5 minutes.
7. Observe the color of the dye.

Applying Mordant
1. Treat the four pieces of clothes of each kind on the bought mordant and soaked it for about 30 minutes in a clean container. This stables the dye on the cloths.

Dyeing Cloth with Round Turmeric
1. Soak the cloths on the container with the dye on it including the four cloths which were applied with mordant.
2. Place the cloths and dyestuffs on the cleaned pyrex again.
3. Boil. Wait for about 15 minutes, and then let it cool for a while.
4. Remove the cloths out from the boiled pyrex then let them dry.

Quality and Discoloration Test
1. Cut all the cloths in halves to have 4 fabrics in each kind of cloth.
2. Discoloration test was performed on the other three cloths.
a. The first cloth was bleached with detergent for one hour.
b. The second was washed with running water for 30 seconds.
c. The third was in sun exposure for 30 minutes.
QUALITY TEST DISCOLORATION TEST
1 – Extreme change in color from white 1- No change in color
2- Change in color from white 2- Less change in color
3- Less change in color from white 3- Change in color
4- No change in color from white 4- Extreme change in color
d. And the fourth was the control or the original fabric with no signs of discoloration.

3. The three fabrics were then compared to the fourth cloth.
4. There were 30 female respondents who compared and graded the dye colorfastness and discoloration qualities. The grading was based as follows:
5. The cloth on its original color will be compared on each other; gina earth, cotton, satin and polyester.
6. The cloths that went to several exposures will also be compared on each other.


CHAPTER IV RESULTS AND DISCUSSION
 There were 30 female respondents who rate the quality of the dye stuff s to come up with these results.

Table A1: Quality of the dye on the fabrics using Round Turmeric
Respondents # Gina Earth Satin Polyester Cotton
1 3 1 2 3 2 3 3 2 1 3 2 2
2 2 1 2 1 3 2 3 1 2 2 3 1
3 2 1 3 1 3 2 2 3 2 2 3 1
4 2 1 2 2 3 3 2 3 1 2 3 1
5 2 1 2 2 3 3 2 3 1 2 3 1
6 2 1 2 2 3 3 2 3 1 2 3 1
7 2 1 2 2 2 3 2 3 1 2 3 1
8 1 1 2 2 2 2 2 3 1 2 2 1
9 2 1 2 2 3 2 2 2 1 2 3 1
10 2 1 2 1 2 2 2 3 1 2 3 1
11 2 1 1 2 1 3 2 3 2 1 3 1
12 3 2 2 3 2 4 3 3 3 3 2 2
13 2 1 2 2 3 3 3 2 1 2 3 1
14 2 1 1 2 2 2 2 3 1 3 3 1
15 1 2 2 3 2 3 1 3 2 1 3 3
16 1 2 2 2 3 2 3 3 2 3 2 2
17 1 2 2 3 2 2 3 2 2 3 2 1
18 1 2 3 2 2 2 3 2 1 2 3 2
19 1 1 2 3 3 2 2 3 1 1 2 2
20 1 1 2 1 2 2 2 4 1 1 2 2
21 1 2 1 1 2 4 2 3 1 2 3 1
22 2 1 2 1 1 3 2 3 2 2 3 1
23 2 1 2 2 1 3 2 3 1 2 3 2
24 1 2 1 1 2 3 1 2 1 2 3 2
25 2 1 2 2 1 2 2 3 2 2 3 1
26 2 1 2 2 1 3 2 3 1 2 3 1
27 2 1 2 2 1 3 2 3 1 1 3 1
28 2 1 1 2 1 3 3 3 1 2 3 1
29 2 1 1 2 3 3 3 3 1 2 2 1
30 2 1 2 2 2 3 2 1 1 2 3 1


QUALITY TEST
1 – Extreme change in color from white
2- Change in color from white
3- Less change in color from white
4- No change in color from white
Legend:


Table A2: Quality of the dye on the fabrics using Mangosteen
1 2 3 1 2 3 3 2 2 3 2 1 3
2 3 2 2 1 3 2 1 3 1 2 1 2
3 3 2 2 3 2 2 3 3 1 3 1 2
4 3 2 1 3 2 3 1 3 2 2 1 2
5 3 2 1 3 2 3 2 3 2 2 1 2
6 3 2 1 3 2 3 2 3 2 2 1 2
7 3 2 1 3 2 3 2 2 2 2 1 2
8 2 2 1 3 2 2 3 2 2 2 1 1
9 3 2 1 2 2 2 2 3 2 2 1 2
10 3 2 1 3 2 2 3 2 1 2 1 2
11 3 1 2 3 2 3 2 1 2 1 1 2
12 2 3 3 3 3 4 1 2 3 2 2 3
13 3 2 1 2 3 3 1 3 2 2 1 2
14 3 3 1 3 2 2 1 2 2 1 1 2
15 3 1 2 3 1 3 1 2 3 2 2 1
16 2 3 2 3 3 2 3 3 2 2 2 1
17 2 3 2 2 3 2 2 2 3 2 2 1
18 3 2 1 2 3 2 2 2 2 3 2 1
19 2 1 1 3 2 2 3 3 3 2 1 1
20 2 1 1 4 2 2 3 2 1 2 1 1
21 3 2 1 3 2 4 3 2 1 1 2 1
22 3 2 2 3 2 3 2 1 1 2 1 2
23 3 2 1 3 2 3 2 1 2 2 1 2
24 3 2 1 2 1 3 2 2 1 1 2 1
25 3 2 2 3 2 2 1 1 2 2 1 2
26 3 2 1 3 2 3 2 1 2 2 1 2
27 3 1 1 3 2 3 3 1 2 2 1 2
28 3 2 1 3 3 3 2 1 2 1 1 2
29 2 2 1 3 3 3 1 3 2 1 1 2
30 3 2 1 1 2 3 2 2 2 2 1 2
Respondents # Gina Earth Satin Polyester Cotton


QUALITY TEST
1 – Extreme change in color from white
2- Change in color from white
3- Less change in color from white
4- No change in color from white
Legend:

• STATISTICAL CALCULATIONS FOR QUALITY TEST
Table 1. Results for Quality Test
Type of Dye Type of Cloth Total
Round Turmeric Dye Satin Gina Earth Polyester Cotton
8.02
2.31 1.91 1.85 1.95
Mangosteen 2.53 2.02 2.0 1.62 8.17
Total 4.84 3.93 3.85 3.57 16.19
a. H0 : There is significant difference on the quality test between the type of cloth (satin, gina earth, polyester, and cotton) dyed in Round Turmeric and Mangosteen.
b. H1 : There is no significant difference on the quality test between the type of cloth (satin, gina earth, polyester, and cotton) dyed in Round Turmeric and Mangosteen.
c. x2 > 7.815 ; Mean = 30
d. Use 0.05 level of significance
e. Computation:
For 2.31: (8.02)(4.84) = 2.44 For 2.53: (8.17)(4.84) = 2.44
16.19 16.19
For 1.91: (8.02)(3.93) = 1.98 For 2.02: (8.17)(3.93) = 1.98
16.19 16.19
For 1.85: (8.02)(3.85) = 1.91 For 2.0: (8.17)(3.85) = 1.94
16.19 16.19
For 1.95: (8.02)(3.57) = 1.77 For 1.62: (8.17)(3.57) = 1.8
16.19 16.19
Table 1.1. Results Using Chi-Square Distribution
OF EF OF-EF (OF-EF)2 (OF-EF)2/EF
2.31 2.44 -0.13 0.0169 0.00693
1.91 1.98 -0.07 0.0049 0.002475
1.85 1.91 -0.06 0.0036 0.001885
1.95 1.77 0.18 0.0324 0.01831
2.53 2.44 0.09 0.0081 0.00332
2.02 1.98 0.64 0.4096 0.2069
2.0 1.94 0.06 0.0036 0.001856
1.62 1.80 -0.18 0.0324 0.018
x2 = 0.2597
f. Decision:
H0 - Accepted ; since /0.2597/ is less than /7.815/. Therefore there is significant
H1 - Rejected difference on the quality test between the type of cloth (satin, gina earth, polyester, and cotton) dyed in Round Turmeric and Mangosteen
 Three obstacles were applied on the fabrics to determine their discoloration rate. The researchers discussed the following results gathered from the experiment.

Table B1: Discoloration after bleaching using Round Turmeric Dye
Respondents # Gina Earth Satin Polyester Cotton
1 3 4 2 4
2 4 4 4 4
3 4 3 4 3
4 4 4 3 4
5 4 3 2 4
6 4 4 3 4
7 4 4 3 4
8 3 4 3 4
9 3 4 4 3
10 4 4 3 4
11 3 4 3 4
12 4 3 4 4
13 4 4 3 4
14 3 4 3 4
15 3 4 2 4
16 3 4 3 4
17 3 4 3 4
18 4 4 4 4
19 4 4 2 3
20 3 4 4 3
21 4 4 3 4
22 4 4 3 4
23 3 3 3 4
24 4 4 3 3
25 4 4 3 4
26 4 4 3 4
27 4 3 3 4
28 4 4 3 4
29 4 3 3 4
30 4 3 3 4

Legend:
DISCOLORATION TEST
1- No change in color
2- Less change in color
3- Change in color
4- Extreme change in color


Table B2: Discoloration after bleaching using Mangosteen Dye


Respondents #
Gina Earth
Satin
Polyester
Cotton
1 3 3 2 4
2 3 4 2 4
3 3 4 3 4
4 3 4 4 3
5 4 4 3 3
6 4 3 2 3
7 4 3 3 3
8 4 3 3 3
9 4 4 3 4
10 3 3 3 4
11 3 4 3 4
12 3 3 3 4
13 4 3 4 4
14 3 3 2 3
15 3 3 2 4
16 3 3 4 4
17 3 3 3 3
18 4 3 4 4
19 4 4 3 4
20 4 4 3 4
21 4 4 3 4
22 3 3 2 4
23 4 3 2 3
24 4 4 3 4
25 3 3 4 4
26 4 4 4 4
27 3 3 4 4
28 4 4 4 4
29 3 4 4 3
30 4 3 4 3
DISCOLORATION TEST
1- No change in color
2- Less change in color
3- Change in color
4- Extreme change in color

Legend:


• STATISTICAL CALCULATIONS FOR DISCOLORATION TEST
Table B: Discoloration after bleaching


Type of Dye Type of Cloth Total
Round Turmeric Dye Satin Gina Earth Polyester Cotton
14.337
3.77 3.67 3.067 3.83
Mangosteen 3.43 3.5 3.067 3.67 13.667
Total 7.2 7.17 6.134 7.5 28.004
a. H0 : There is significant difference on the discoloration test undergoing bleaching for 30 minutes between the type of cloth (satin, gina earth, polyester, and cotton) dyed in Round Turmeric and Mangosteen.
b. H1 : There is no significant difference on the discoloration test undergoing bleaching for 30 minutes between the type of cloth (satin, gina earth, polyester, and cotton) dyed in Round Turmeric and Mangosteen.
c. x2 > 7.815 ; Mean = 30
d. Use 0.05 level of significance
e. Computation:

For 3.77: (14.337)(7.2) = 3.686 For 3.43: (13.667)(7.2) = 3.514
28.04 28.004


For 3.67: (14.337)(7.17) = 3.671 For 3.5: (13.667)(7.17) = 3.499
28.004 28.004

For 3.067: (14.337)(6.134) = 3.140 For 3.067: (13.667)(6.134) = 2.994
28.004 28.004

For 3.83: (14.337)(7.5) = 3.840 For 3.67: (13.667)(7.5) = 3.660
28.004 28.004


Table 2. Results Using Chi-Square Distribution


OF EF OF-EF (OF-EF)2 (OF-EF)2/EF
3.77 3.686 0.084 0.00706 0.001915
3.67 3.671 -0.001 0.000001 0.00000027
3.067 3.140 -0.073 0.00533 0.001697
3.83 3.840 -0.01 0.0001 0.000026
3.43 3.514 -0.084 0.00706 0.00201
3.5 3.499 -0.001 0.000001 0.000000286
3.067 2.994 0.073 .00533 0.00178
3.67 3.660 0.01 0.0001 0.0000273
x2 = 0.0074559

f. Decision:
H0 - Accepted ; since /0.0074559/ is less than /7.815/. Therefore there is significant
H1 - Rejected difference on the discoloration test undergoing bleaching for 30 minutes between the type of cloth (satin, gina earth, polyester, and cotton) dyed in Round Turmeric and Mangosteen
 On the bleaching test Cotton retained the most color among the others as what the respondents see. And as expected, the Satin fabric had the better result than the other clothes.


Table C1: Discoloration after pouring with water using Round Turmeric Dye

Respondents #
Gina Earth
Satin
Polyester
Cotton
1 3 2 3 3
2 2 2 3 3
3 2 2 3 3
4 2 2 2 3
5 2 2 2 3
6 3 2 2 3
7 2 2 2 3
8 2 2 2 3
9 2 2 2 3
10 2 2 2 3
11 1 2 2 2
12 1 2 2 3
13 3 2 2 2
14 2 2 2 3
15 2 2 2 3
16 2 2 2 3
17 3 1 2 3
18 3 1 2 2
19 3 2 1 3
20 3 2 2 3
21 1 2 2 3
22 2 2 2 3
23 2 2 2 3
24 2 3 2 3
25 2 2 2 4
26 2 2 2 3
27 1 2 2 3
28 2 2 2 3
29 3 2 2 3
30 2 2 2 3

Legend:
DISCOLORATION TEST
1- No change in color
2- Less change in color
3- Change in color
4- Extreme change in color


Table C2: Discoloration after pouring with water using Mangosteen Dye

Respondents #
Gina Earth
Satin
Polyester
Cotton
1 3 2 1 3
2 2 2 2 3
3 2 2 2 3
4 2 2 2 3
5 2 2 2 2
6 2 3 3 3
7 2 2 2 2
8 2 3 3 2
9 2 1 3 3
10 1 1 2 3
11 2 2 2 3
12 2 2 2 3
13 1 2 2 2
14 3 2 2 2
15 3 2 3 2
16 3 3 2 2
17 2 2 3 3
18 2 2 1 2
19 2 2 3 3
20 2 2 1 3
21 2 2 3 4
22 2 2 3 3
23 2 2 3 3
24 3 2 2 3
25 1 2 2 3
26 2 2 2 3
27 2 2 2 3
28 2 3 2 3
29 2 2 2 3
30 3 2 2 3

Legend:
DISCOLORATION TEST
1- No change in color
2- Less change in color
3- Change in color
4- Extreme change in color


• STATISTICAL CALCULATIONS FOR DISCOLORATION TEST
Table C: Discoloration after pouring with water
Type of Dye Type of Cloth Total
Round Turmeric Dye Satin Gina Earth Polyester Cotton
9.06
1.93 2.13 2.07 2.93
Mangosteen 2.03 2.1 2.2 2.77 9.1
Total 3.96 4.23 4.27 5.7 18.16
a. H0 : There is significant difference on the discoloration test undergoing pouring with water for 30 seconds between the type of cloth (satin, gina earth, polyester, and cotton) dyed in Round Turmeric and Mangosteen.
b. H1 : There is no significant difference on the discoloration test undergoing pouring with water for 30 seconds between the type of cloth (satin, gina earth, polyester, and cotton) dyed in Round Turmeric and Mangosteen.
c. x2 > 7.815 ; Mean = 30
d. Use 0.05 level of significance
e. Computation:
For 1.93: (9.06)(3.96) = 1.98 For 2.03: (9.1)(3.96) = 1.98
18.16 18.16

For 2.13: (9.06)(4.23) = 2.11 For 2.1: (9.1)(4.23) = 2.12
18.16 18.16

For 2.07: (9.06)(4.27) = 2.13 For 2.2: (9.1)(4.27) = 2.14
18.16 18.16

For 2.93: (9.06)(5.7) = 2.85 For 2.77: (9.1)(5.7) = 2.86
18.16 18.16



Table 3. Results Using Chi-Square Distribution

OF EF OF-EF (OF-EF)2 (OF-EF)2/EF
1.93 1.98 -0.05 0.0025 0.001263
2.13 2.11 0.02 0.0004 0.0001896
2.07 2.13 -0.06 0.0036 0.00169
2.93 2.85 0.08 0.0064 0.00225
2.03 1.98 0.05 0.0025 0.001263
2.1 2.12 -0.02 0.0004 0.0001887
2.2 2.14 0.06 0.0036 0.00168
2.77 2.86 -0.09 0.0081 0.002832
x2 = 0.011356
f. Decision:
H0 - Accepted ; since /0.011356/ is less than /7.815/. Therefore there is significant
H1 - Rejected difference on the discoloration test undergoing pouring with water for 30 seconds between the type of cloth (satin, gina earth, polyester, and cotton) dyed in Round Turmeric and Mangosteen.
 On the rubbing through running water, as what we've expected, the cotton fabric and the Gina earth again retained the most color. Surprisingly the Polyester and satin fabrics had the lowest average on retaining the color which means they only retain a little pigment.

Table D1: Results after Cloths underwent sunlight exposure for 30 minutes using Round Turmeric Dye



Respondents #
Gina Earth
Satin
Polyester
Cotton
1 2 3 2 3
2 2 3 2 2
3 3 2 2 2
4 2 3 3 2
5 2 3 3 2
6 3 3 2 2
7 3 3 2 2
8 3 3 2 2
9 3 3 2 2
10 3 3 2 2
11 3 3 2 2
12 2 2 3 3
13 3 3 2 2
14 2 2 2 2
15 2 3 2 3
16 2 1 3 3
17 2 3 2 2
18 2 3 2 2
19 3 2 3 3
20 2 2 4 3
21 3 3 2 2
22 3 3 2 1
23 2 3 2 2
24 3 3 2 2
25 3 3 2 1
26 3 3 2 2
27 3 3 2 2
28 3 3 2 2
29 2 3 2 2
30 3 3 2 2


Legend:
DISCOLORATION TEST
1- No change in color
2- Less change in color
3- Change in color
4- Extreme change in color


Table D2: Results after Cloths underwent sunlight exposure for 30 minutes using Mangosteen Dye


Respondents #
Gina Earth
Satin
Polyester
Cotton
1 3 3 2 2
2 3 3 2 2
3 3 3 3 2
4 3 2 3 2
5 3 2 3 2
6 3 2 2 2
7 3 2 2 2
8 3 2 2 2
9 3 2 2 2
10 3 2 2 2
11 2 2 3 2
12 3 2 3 2
13 2 2 3 2
14 3 2 3 2
15 3 2 2 2
16 3 2 2 2
17 3 2 2 1
18 2 2 4 1
19 3 1 2 2
20 3 2 2 2
21 3 2 2 2
22 3 2 2 2
23 3 2 3 2
24 3 2 2 3
25 4 2 2 2
26 3 2 3 2
27 3 2 3 2
28 3 2 2 2
29 3 2 2 2
30 3 2 2 2

Legend:
DISCOLORATION TEST
1- No change in color
2- Less change in color
3- Change in color
4- Extreme change in color

Table D: Results after Cloths underwent sunlight exposure for 30 minutes



Type of Dye Type of Cloth Total
Round Turmeric Dye Satin Gina Earth Polyester Cotton 9.7

2.77 2.57 2.23 2.13
Mangosteen 2.067 2.93 2.4 1.97 9.367
Total 4.837 5.5 4.63 4.1 19.067
a. H0 : There is significant difference on the discoloration test undergoing sunlight exposure for 30 minutes between the type of cloth (satin, gina earth, polyester, and cotton) dyed in Round Turmeric and Mangosteen.
b. H1 : There is no significant difference on the discoloration test undergoing sunlight exposure for 30 minutes between the type of cloth (satin, gina earth, polyester, and cotton) dyed in Round Turmeric and Mangosteen.
c. x2 > 7.815 ; Mean = 30
d. Use 0.05 level of significance
e. Computation:
For 2.77: (9.7)(4.837) = 2.461 For 2..067: (9.367)(4.837) = 2.376
19.067 19.067

For 2.57: (9.7)(5.5) = 2.798 For 2.93: (9.367)(5.5) = 2.702
19.067 19.067

For 2.23: (9.7)(4.63) = 2.355 For 2.4: (9.067)(4.63) = 2.275
19.067 19.067

For 2.13: (9.7)(4.1) = 2.086 For 1.97: (9.067)(4.1) = 2.014
19.067 19.067

Table 3. Results Using Chi-Square Distribution


OF EF OF-EF (OF-EF)2 (OF-EF)2/EF
2.77 2.461 0.309 0.095481 0.038798
2.57 2.798 -0.228 0.051984 0.018579
2.23 2.355 -0.125 0.015625 0.006635
2.13 2.086 0.044 0.401936 0.000929
2.067 2.376 -0.309 0.095481 0.040186
2.93 2.702 0.228 0.051984 0.019239
2.4 2.275 0.125 0.015625 0.006868
1.97 2.014 -0.044 0.001936 0.000961
x2 = 0.13219
f. Decision:
H0 - Accepted ; since /0.132195/ is less than /7.815/. Therefore there is significant
H1 - Rejected difference on the discoloration test undergoing sunlight exposure for 30 minutes between the type of cloth (satin, gina earth, polyester, and cotton) dyed in Round Turmeric and Mangosteen.
 The last obstacle was sunlight exposure for 30 minutes. They hardly retained their color But there are some who retained more and retain less. Cotton and polyester were the fabrics that retained more color while satin fabric retained less.

DISCUSSIONS:
After the experiment the researchers extracted the dye from the Round Turmeric rhizomes. The dye was a little sticky because it was boiled. When it is applied on cloths, the dye which adheres to the fabric was not good compared to the researcher's expectations.

CHAPTER V
CONCLUSIONS AND RECOMMENDATIONS
CONCLUSIONS
The researchers conclude the following:
1.) The researchers with the respondents concluded that the color of the dye is Pale yellow to dark yellow.
2.) After summing up their rates, the cotton fabric was chosen as the cloth which retains dye
color even passing through bleaching, rubbing with running water, and exposure.
3.) The researchers with the respondents concluded that Satin fabric exhibits an attractive and glamorous color among the others.
4.) Based from the results above, the researchers concluded that rhizomes from Round Turmeric have the potential in producing dye out of it.

RECOMMENDATIONS
The researchers would like to recommend the following:
1.) To use other kinds of mordant in order to improve the quality of the dye.
2.) The researchers would also like to recommend different kinds of textiles by using Round Turmeric dye.
3.) Try also to compare the round turmeric dye to commercial ones.
4.) The researchers would also recommend other plants to be used.

5.) If possible, discover another color that round turmeric rhizomes can produced aside from golden and dark yellow.
RESPONDENT'S PAPER
Quality
of dye Gina Earth Satin Polyester Cotton
Color of Yellowish brown
Dye
Discoloration

Bleached
Poured
with
Water
Sunlight Exposure
Name & Signature of Respondent











APPENDICES


THE MATERIALS USED





















WEIGHING OF ROUND TURMERIC



















PEELING OF RHIZOMES






















POUNDING OF RHIZOMES
























BOILING THE POUNDED ROUND TURMERIC























THE CLOTHES SOAKED WITH MORDANT























BOILING THE EIGHT CLOTHES IN THE DYE
























THE DYE WITH THE CLOTHES BOILING ON THE PYREX




















THE DYE STUFF BEING HANGED























THE CLOTHES WHICH WERE DYED























DISCOLORATION TEST

CLOTHES EXPOSED TO SUNLIGHT
 DURING





















 AFTER





















CLOTHES WITH RUNNING WATER
 DURING






















 AFTER























CLOTHES SOAKED WITH DETERGENT
 DURING























 AFTER






















BIBLIOGRAPHY



Internet Sites:

(http://en.wikipedia.org/wiki/Curcumin)

(http://www.mdidea.com/products/new/new088.html#01)

(http://www.evitamins.com/healthnotes.asp?ContentID=2175005)

(http://webexhibits.org/causesofcolors/2C.html)

(http://pubs.acs.org/isubscribe/journals/joceah/65/i21/figures/jo000335zh00002.html)

(http://www.hort.cornell.edu/plantsandtextiles/pdfs/indigo/introduction.pdf)

(http://www.knaw.nl/ecpa/ink/ink_chemical.html)

(http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6T99-4KR3JRN-1&_user=10&_coverDate=10%2F19%2F2006&_rdoc=1&_fmt=&_orig=search&_sort=d&view=c&_acct=C000050221&_version=1&_urlVersion=0&_userid=10&md5=4cd385a5c1de1dcfed135c0fd7b8055b)

(http://www.ncbi.nlm.nih.gov/sites/entrez?cmd=Retrieve&db=PubMed&dopt=AbstractPl us&list uids=17959149)


Books:

“Dye”. Groiler Children’s Encyclopedia. 1994. Vol.1.

“Regional Assessment and Collection of Available Dye-Yielding Plants in the
Philippines” Philippine Technology Journal. Taguig, Metro Manila:STII-DOST.
2000.

Batu, Ruby Ann Celeste. “Extracted Dye from Camote Tops” A Junior Research Paper,
MSU-IIT, IDS, Iligan City (2004)

Tibor, Karyl Hamot. “Dye from Coconut Roots”. MSU-IIT, IDS, Iligan City. (1996)

Castillo, Casharelle. “ Comparative Study on the Colorfastness of Commercially
Availiable Synthetic Dye and Natural Dye from Gemilina”
A Junior Research Paper, MSU-IIT, IDS, Iligan City. (2002)

Lomansoc, Jade Kenneth. “Production of Dye from the Sap of an Avocado Seed”
MSU-IIT, IDS, Iligan City. (2004)

BIOGRAPHICAL DATA

Name : Vanica Irish M. Bungco
Nickname : Van-Van, Nike
Place of Birth : Iligan City
Date of Birth : December 13, 1991
Home Address : Block 20 Lot 7 Steeltown Iligan City
City Address : Region 10, Iligan City
Civil Status : Student
Sex : Female
Religion : Roman Catholic
------------------------------------------------------------------------------------------------------------------


BIOGRAPHICAL DATA

Name : Karen B. Buhia
Nickname : Karz
Place of Birth : Iligan City
Date of Birth : January 21, 1991
Home Address : Bagong Silang, Iligan City
City Address : Region 10, Iligan City
Civil Status : Student
Sex : Female
Religion : Seventh-Day Adventist
BIOGRAPHICAL DATA

Name : April I. Paradela
Nickname : Pril
Place of Birth : Iligan City
Date of Birth : April 13, 1992
Home Address : Buru-un, Iligan City
City Address : Region 10, Iligan City
Civil Status : Student
Sex : Female
Religion : Roman Catholic
------------------------------------------------------------------------------------------------------------------


BIOGRAPHICAL DATA

Name : Michelle G. Sarona
Nickname : Mitch
Place of Birth : Iligan City
Date of Birth : September 24, 1991
Home Address : Ditucalan, Iligan City
City Address : Region 10, Iligan City
Civil Status : Student
Sex : Female
Religion : Roman Catholic

BIOGRAPHICAL DATA

Name : Desimie Apple L. Gumabat
Nickname : Dez
Place of Birth : Iligan City
Date of Birth : April 7, 1992
Home Address : Tubod, Iligan City
City Address : Region 10, Iligan City
Civil Status : Student
Sex : Female
Religion : Christian


























“DYE FROM ROUND TURMERIC RHIZOMES”


A Research Paper


Presented to
Iligan City National High School
Iligan City


In Partial Fulfillment of the Requirements for the
Requirements for Research II

Submitted by:
Vanica Irish Bungco
Michelle G. Sarona
April Rose Paradela
Desimie Gumabat
Karen Buhia



Submitted to:
Mrs. Annallee Aron
(Research Adviser)




APPROVAL SHEET


This research paper attached here, entitled “Dye from Round Turmeric Rhizomes” prepared and Vanica Irish Bungco, April Rose Paradela, and Karen Buhia, Michelle G. Sarona, and Desimie Apple L. Gumabat in partial fulfillment of Research II, is recommended for approval.


Aimee Pareño Alberto Cabungcal Annallee Aron
Panel Member Panel Member Panel Member


This Research paper is hereby approved in fulfillment of the required for the course Research.

Annallee Aron
Research II Adviser

Nida H. Gumera
Science Department Head

Cytoxicity of Lemon Grass (Cymbopogon citratus) Extract

Cytoxicity of Lemon Grass (Cymbopogon citratus) Extract
Though Brine Shrimp Bioassay



A Research Paper
Presented to the
Department of Science and Technology
(Special Science Curriculum)
Iligan City National High School
Iligan City

__________________

In Fulfillment of the Requirements
In Research II
_________________

Mrs. Annallee Q. Aron
Adviser
__________________


By:
Adanza, Faith Stephanie E.
Bala, Clendelyn Jane D.
Redublado, Marjorie S.


March , 2008

ABSTRACT
Nowadays, there are many plants found in our environment, some of it is used in our daily living such as spices. But yet, we don’t know if these plants are bioactive. Determining plants bioactivity can lead to discoveries of the other uses of this plant, especially in the field of medicine.
This research study aims to determine the bioactivity of lemon grass extract through brine shrimp bioassay.
Lemon grass is commonly use as spices in the Philippines and is also known in our dialect us “Tanglad”.
Brine shrimp bioassay uses brine shrimps to be able to determine how bioactive a certain plant is.
The brine shrimps are hatched for two days and after that, preparations of the test solutions are made. The researchers prepared three test-solutions namely 1000-ppm, 100-ppm, 10-ppm. After getting the number of mortality on different concentrations and the data is tabulated in table form and in a plotted in a graph. The researchers use ANOVA statistical test in determining the significant difference between the experimental set-up and the control. The researchers also use the table in finding the LC50 of experimental set-up.
Results from ANOVA statistical test shows that there is a significant difference between the experimental set-up and the control. Results from the LC50 table shows that the number of concentration is directly proportional to the number of mortality, and LC50=52.596.
From the result of the ANOVA statistical test and LC50, it was then concluded that lemon grass is bioactive.

APPROVAL SHEET

The research proposal entitled “Cytoxicity of Lemon Grass Extract Though Brine Shrimp Bioassay”, prepared by Clendelyn Jane D. Bala, Marjorie S. Redublado, Faith Stephanie Adanza, in fulfillment of the requirements in Research II have been examined and recommended for acceptance and approval.

Annallee Q. Aron
Research II Adviser



Marjorie Y. Dumaug Aimee R. Pareño Annalle Q. Aron
Panel Member Panel Member Panel Member

This research proposal is hereby approved in fulfillment of the requirements for the subject Research II.

Nida H. Gumera
Science Dept. Head (Day)


ACKNOWLEDGEMENT

The researchers express their thanks and sincere praises to the Most Merciful and Most Loving God for the guidance and strength which they needed to overcome all the obstacles in their life. Without God’s help and love, this research work would have not been possible.

They also express their thanks and sincere gratitude to the following whose contribution have made the completion of this research paper:
Mrs. Annallee Aron, their research adviser, whose intellectual guidance and sound advices made possible to complete this research paper.
Ms. Victoria Cordova, for letting them use the computer laboratory for the completion of their research.
Panel members, whose comments and suggestions made changes in some parts of this research paper.
Ms. Desimie Gumabat, for allowing the researcher to continue their unfinished study, together with her partner.
To the brine shrimp team (Anamel, Alyzza and Ginuel) for the sleepless nights they’ve shared together. For the friendship and conversations they have been through.
There classmates, Maxtein ’08, for the laughter, kindness and companionship throughout the high school years.
Finally, the researchers express their deepest gratitude to their family for their support (financially and emotionally), enduring love, encouragement, and understanding to finish the research paper.
To all persons who have contributed to this paper, thank you very much.

Clenz, Faith, Marj

TABLE OF CONTENT

Title Page ………………………………………..………………………
Abstract ………………………………………..………………………
Approval Sheet ………………………………..………………………
Acknowledgement ………………………………..………………………
Table of Contents …………………………………………..……………

CHAPTER I- Introduction
Background of the Study …………………………………..……
Statement of the Problem …………………………………..……
Significance of the Study ……………………………..…………
Scope and Delimitation ……………………………..…………

CHAPTER II – Review of Related Literature and Studies
Review of Related Study ………………………………………..
Conceptual Framework ………………………………………..
Null Hypothesis ………………………………………..
Definition of Terms ………………………………………..

CHAPTER III – Methodology
Materials ………………………………………..
Procedure ………………………………………..
Data Gathering Procedure ………………………………………..
Statistical Used ………………………………………..
Instruments Used ………………………………………..
CHAPTER IV- Results and Discussions
Table1 ……………………………………………………..…
Chart 1 ……………………………………………………..…

Table 2 ……………………………………………………..…
Chart 2 ……………………………………………………..…
Decision ……………………………………………………..…
CHAPTER IV – Summary, Conclusion and Recommendation
Summary ……………………………………………………..…
Conclusion ……………………………………………………..…
Recommendation ……………………………………………..…
BIBLIOGRAPHY ……………………………………………..…
APPENDICES ……………………………………………………..…

CHAPTER I
INTRODUCTION

Background of the Study

Tanglad or Lemongrass extract (Cymbopogon citratus) s a fragrant grass that grows in abundant clumps just about anywhere in the tropics. It possesses a larger bulb near the base stalks and it is here that the lemon or citrus flavoring is most often obtained ( though we Filipinos use the entire leaf for stuffing ).Lemongrass is used as a base for curries, soups, stews and it is also used to flavor salads, as part of marinades and as stuffing material. It can be infused into hot water for a simple herbal infusion and it now flavors desserts as well (http:/www.marketmanila.com/archives/tanglad-lemongrass).
Just over a centimeter in size, the adult brine shrimp Artemia is an extremely well-known animal because of its importance as a food source for fish and crustaceans raised in home aquariums, aquaculture systems and in laboratories. One can buy brine shrimp at practically any pet display. It looks like a powdery brown substance but in reality the substance is thousands of cysts—eggs surrounded by protective cases. When added to water, these cysts will hatch into shrimp nauplii within a few hours (http://seagrant.gso.uri.edu/factsheets/931brine.html).
Brine shrimp are food source not only for humans but also for aquatic life, since they are found naturally in about 250 locations around the world. Brine shrimp have been used as a “bench top bioassay” for the discovery and purification of bioactive natural products and they are an excellent choice for elementary toxicity investigations of consumer products (http://scholar.google.com/q=importance of brine shrimp bioassay=scholart).
Bioassay is shorthand commonly used term for biological assay and is a type of scientific experiment. Bioassays are typically conducted to measure the effects of a substance on a living organism. It may be qualitative or quantitative. Qualitative bioassays are used for assessing the physical effects of a substance that may not be quantified, such as abnormal development or deformity. Quantitative bioassays involve estimation of the concentration or potency of a substance by measurement of the biological response that it produces. Quantitative bioassays are typically analyzed using the methods of biostatistics. Bioassays are essential in the development of new drugs and monitoring pollutants in the environment (Microsoft Corporation © 1993-2003).

Statement of the Problem
This study aims to answer the following questions:
1.) Is there any significant difference in terms of brine shrimp lethality with the lemongrass extract and its control?
2.) Is there any significant difference in terms of toxicity in different concentrations?

Significance of the Study
This research study is about identifying toxicity of Lemongrass (Cymbopogon citratus) extract through Brine Shrimp Bioassay having different concentrations.
Brine shrimp have been used as “bench top bioassay” for the discovery and purification of bioactive natural products and they are excellent choice for elementary toxicity investigations of consumer products.
This could help future researches who will be conducting studies relating to lemongrass to decide if lemongrass would fit to their study.

Scope and Delimitation
This research study was conducted at Iligan City National High School, ESEP building last February 23, 2008. The focus of the study is to test the toxicity of lemongrass extract in order to find out how bioactive lemongrass is. The extraction was conducted at MSU-IIT (Chemistry Laboratory of College, Science and Mathematics) last December 20, 2007.

CHAPTER II
REVIEW OF RELATED STUDY AND LITERATURE

Related Study
“A brine Shrimp Bioassay for measuring Toxicity and Remediation of Chemicals”
This article describes a laboratory project that uses a bioassay on brine shrimp to measure LC50 values or a variety of insecticides. This project was the center piece of a five-week unit on toxicology and chemical remediation. The 22 students in the class were first-year. Two 3-hour lab periods are required, the first for toxicity testing, and the second for remediation and retesting. Any chemical that can be dissolved or dispersed into water can be studied in this bioassay; plant extracts or other natural products. Although the bioassay is simple enough to use with student who have had little lab experience, it could also be integrated into an environmental, organic, medical or biochemistry lab course.
This set of labs helps student how toxicity is measured and how harmful molecules can be decomposed to harmless products.
The bioassay and remediation procedures described in the supplementary material could be used as straight and remediation procedures described in the supplementary material could be used as straight labs as a multi-week project. In such project, students were given a letter from the owner of a dog-grooming salon, who was worried that the insecticide products she uses in the salon are leaching into a nearby stream. She asks the students to test the flea products to determine their toxicity in aquatic life, and to devise some method that could use to make the wastewater less toxic before she pours it down the drain.
The students were then divided into groups of 3 or 4 and given an insecticide product. They measured the LC50 value of their insecticide o brine shrimp; all seven groups were able to complete this lab in three-hour lab period. Over the next two-weeks, the groups researched the chemicals present in their insecticide product. In the second lab period, the remediation procedures were carried out and the remediate material was assayed on brine shrimp (Marya.Lieberman, 1994).

Conceptual Framework
Independent Variable Dependent Variable

Null Hypothesis
1. There is no significant difference in terms of brine shrimp lethality with the lemongrass extract and its control.
2. There is no significant difference in terms of toxicity in different concentration.

Definition of Terms
1. Artificial Seawater –can be attained by dissolving 40g of salt in 100ml of water. Also used as substitute for pure seawater
2. Brine Shrimp –very tiny crustaceans that are easy to breed and maintain for long period of time and are commonly used as food for marine fish
3. Bioassay –a test used to evaluate the relative potency of a chemical or mixture of chemicals by comparing its effect on a living with the effect of a standard preparation on the same organism
4. Compressor –a mechanical devise that increases the pressure of gas
5. Cyst –are sac-like lesion containing liquid material consisting of white blood cells, dead cells, and bacteria
6. Extract –to obtain from a substance by chemical or mechanical action, as by pressure, distillation, or evaporation
7. Lemongrass -is a genus of about 55 species of grasses, native to warm temperate and tropical regions of the old world and Ocennia. It is a tall perennial grass; common names include lemongrass, lemon grass, barbed wire grass, silky heads citronella grass, fever grass or Hierba Luisa
8. Ppm –a unit of measure used to express the number of parts of substance contained within a million parts of a liquid, solid, or gas
9. Nauplii –a newly hatched brine shrimp
10. Yeast -any of a number of microscopic, one-celled fungi important for their ability to ferment carbohydrates in various substances

CHAPTER III
METHODOLOGY

Materials
20 Test tubes Lemon grass
Jar Brine Shrimp
Syringe Yeast
500mL Distilled water Graduated Cylinder
20 grams Salt Paper Disc

Procedure
I. Gathering and of Lemon grass
Lemon grasses were gathered in Tambo, Iligan City.
II. Preparation of Lemon grass Extract
The gathered lemon grass was cut for about ½ inch. Then, it was air-dried for about 3 hours. Then, the air-dried lemongrass was soaked to 900mL of ethanol. Then the soaked lemon grass were sent to MSU-IIT College of Science and Mathematics for the extraction.
III. Brine Shrimp Assay (BSA)
The brine shrimp assay (BSA) was employed to determine the toxicity and general bioactivity of lemon grass.
a. Hatching of Brine Shrimp
Brine shrimp eggs were made to hatch into a jar. The jar is divided into two compartments; half of the compartment is wrapped around to prevent light. Into this jar, artificial sea water is prepared by dissolving 20grams of ordinary table salt in 500mL distilled water. Brine shrimp cysts are then placed and allowed to hatch. The tank should not be moved after adding the eggs to prevent the eggs from getting to other compartment. The sea water is supplemented with yeast to serve as food for the shrimp. After 48 hours of incubation at room temperature, the active brine shrimp nauplii are ready for transfer to the test solution.
b. Preparation of Test Solution of Lemongrass Extract
i. Prepare paper disc of about 0.6cm in diameter.
ii. Calibrate twenty (20) test tubes to 5.0mL volume with a 5.0-mL disposable syringe.
iii. Prepare a stock solution of 50,000-ppm by dissolving 50grams of lemongrass extract in 1.0mL of the artificial seawater.
iv. First, prepare test-tubes of 1,000-ppm test solutions. Transfer 0.1mL of the 50,000-ppm stock solution to each of four test tubes (label a1, a2, a3, a4). Test tube a1, a2, a3 should contain the prepared paper disc and was not air dried.
v. Next, prepare test-tubes of 100-ppm test solutions. Transfer 0.5mL of the 1,000-ppm stock solution to each of four test tubes (label b1, b2, b3, b4). Test tube b1, b2, b3 should contain the prepared paper disc and was not air dried.
vi. Then, prepare test-tubes of 10-ppm test solutions. Transfer 0.5mL of the 100-ppm stock solution to each of four test tubes (label c1, c2, c3, c4). Test tube c1, c2, c3 should contain the prepared paper disc and was not air dried.
IV. Bioassay
Add about 5ml of seawater to each of the test tubes to air-dried extract. Using a syringe, ten (10) brine shrimp nauplii are transferred with seawater from the lighted compartment of the tank into each of the test tubes containing the air-dried samples. Then, the volumes of the test solutions are adjusted to 5ml with artificial seawater.
Data Gathering Procedure
After 24 hours, the survivors are counted and the number of mortality is recorded for each test solution and reported as the average of three replicates. The percent mortality in test solution is calculated as:
Number of mortality
% mortality = ----------------------------- x 100
Number of test shrimp

Statistical Test Used
The statistical test used in this study is ANOVA.

Instruments Used
A. Compressor

CHAPTER IV
RESULTS AND DISCUSSION
In this chapter, discussion about the results will be the focus of the research showing the bioactivity of “Lemongrass” through brine shrimp bioassay.

Table 1
Concentration Test tube 1 Test tube 2 Test tube 3 Average
1000-ppm 9 8 10 9
100-ppm 6 8 7 7
10-ppm 4 2 5 3.7
Ethanol 10 10 10 10
A. Seawater 0 0 0 0
The table shows the number of the brine shrimp mortality in different concentration. It is shown here that 1000-ppm concentration have the highest number of mortality in terms of the three set-up.
Chart 1

Ld50= 52.596
The chart shows the comparison of mortality of different test tubes in different concentration. It shows that the higher the concentration, the higher the mortality. And base on ld50 result, it shows that the experiment shows bioactivity.
Table 2
Concentration Test tube 1 Test tube 2 Test tube 3 Average
1000-ppm 90% 80% 100% 90%
100-ppm 60% 80% 70% 70%
10-ppm 40% 20% 50% 37%
Ethanol 100% 100% 100% 100%
A. Seawater 0% 0% 0% 0%
The table shows the percentage of the brine shrimp mortality in different concentration. It is shown here that 1000-ppm concentration have the highest number of mortality in terms of the three set-up.
Chart 2

Ld50= 50.576
The chart shows the comparison of mortality of different test tubes in different concentration in percentage. It shows that the higher the concentration, the higher the mortality. And base on ld50 result, it shows that the experiment shows bioactivity.

Anova: Two-Factor Replication

SUMMARY Count Sum Average Variance
Row 1 4 36 9 0.666667
Row 2 4 28 7 0.666667
Row 3 4 14.7 3.675 1.555833
Row 4 4 40 10 0

Column 1 4 29 7.25 7.583333
Column 2 4 28 7 12
Column 3 4 32 8 6
Column 4 4 29.7 7.425 7.7225
Decision
ANOVA
Source of Variation SS df MS F P-value F crit
1.15E-
Rows 93.41688 3 31.13896 43.11134 0.5 3.862548
Columns 2.166875 3 0.722292 1 0.4362 3.862548
Error 6.500625 9 0.22292

Total 102.0844 15

Base on the statistical test ANOVA, it shows that Ho is rejected. Therefore H1 is accepted, it is because there is significant difference in terms of mortality of brine shrimp in the experimental and control. And there is also significant difference of mortality in different con concentration.

The researchers have observed that as the concentration of the crude extract of lemon grass increases, the number of mortality also increases.

CHAPTER V
SUMMARY, CONCLUSION, RECOMMENDATION

Summary
This study was conducted at Iligan City National High School, ESEP building, last February 23, 2008 It is all about finding out the bioactivity of lemongrass extract though brine shrimp bioassay. After conducting the bioassay, it was found out that lemongrass has bioactive compound but its bioactivity depends upon the concentrations of the test solution.
The table for mortality of brine shrimp extract shows that lemongrass is more bioactive at higher concentration.

Conclusion
Based on the LC50 and ANOVA statistical test, it shows that there is significant difference between the control and the experimental set-up with different concentrations.
Therefore it was concluded that lemon grass is more bioactive. And the concentration of the lemon grass extract is directly proportional to the mortality of the brine shrimps.

Recommendation
It is highly recommended by the researchers that lemon grass can be used as a raw material for making product that can be used in studies involving bioactivities.
The researchers also recommend the future researchers to use lemongrass in making product such as anti-fungal ointment.
It is also suggested to the future researchers to conduct other form of bioassay to further determine the bioactivity of lemongrass and to conduct study in other different concentration.

BIBLIOGRAPHY

Books
Tim Johnson, CRC Ethanobotany Desk Reference, N.Y. New York, Newday Publishing House, 1993, p.193.

Dictionary
Webster’s Third New International Dictionary, 1995, page 633.

Encyclopedia (Online)
“Bioassay”, Encarta Encyclopedia, Microsoft Corporation, 1993-2003, all rights reserved.

Internet
http://www.seagrant.gso.uri.edu
http://www.scholar.google.com
http://www.sciencedirect.com
http://www.marketmanila.com
http://www.nd.edu

Acacia Bark as Dye


__________________

A Research Proposal Represented to the Department of Science and Technology
(Special Science Curriculum)
Iligan City National High School
Iligan City

____________________

Submitted to:
Mrs. Annallee Q. Aron
Research Adviser
____________________

Submitted by:
Carmille Soliel C. Tabanao
Darlyn E. Tabañag
Catherine Grace N. Anggo

S.Y. 2007-2008

Approval Sheet

A research proposal entitled “Acacia Bark as Dye “ is proposed and submitted by Carmille Soliel C. Tabanao, Darlyn E. Tabañag and Catherine Grace N. Anggo of Einstein in fulfillment in Research IV has been examined for acceptance and approval.

Mr. Alberto S. Cabungcal Ms. Aimee R. Pareño Ms. Marjorie Y. Dumaug
Panel Member Panel Member Panel Member

The research is hereby approved in fulfillment in Research IV

Mrs. Annallee Aron Nida H. Gumera
Research Adviser Science Department Head
(1st and 2nd shifts )

i
Acknowledgement

This research paper has been made possible through the valuable contributions of various people to whom grateful acknowledgement is hereby made.
First of all the researchers would like to thank their parents for their precious encouragement, understanding, and their solicitous concern throughout the whole work. As well as for their financial and moral support which have helped the researchers a lot.
To Mrs. Annallee Q. Aron, their Research II Adviser, for her undying assistance and suggestion which have resulted to the completion and success of this research paper.
To the panelist members, thank you for sharing your knowledge to improved the Researchers study and as well as to your ideas and opinions to the research paper.
To the Researchers friends and classmates who have shared some knowledge and for the inspiration and full support for the success of the research paper.
And most of all to our Father Almighty for his guidance and for giving the researchers the knowledge and strength they need to complete this research paper.
ii

Abstract

The Acacia Bark is usually in curved pieces, externally grayish brown, darkening with age, often with irregular longitudinal ridges and sometimes transverse cracks. Acacia bark contains from 24 to 42 percent of tannin and also Gallic acid. The rampant production of dye can result to the declination of dye prices in markets which provokes lower quality of dye products. Because of these constituents and problems met the researcher are inspired to produce a dye from it
Acacia barks were gathered and were air-dried. Then soaked into the beaker with an ethanol and boiled to get the extract. Lastly, the different types of fabrics were submerged and dried for testing.
Based on the data gathered through a survey, acacia bark extract is good and can be a high-quality dye but its efficiency is greatly showed when using thick fabrics. The intense black color of dye, from Acacia bark, show that it has a possibility to be use as an alternative source of an Ink.


iii
Table of Contents

Page
Title Page
Approval Sheet ………………………………………………………….. i
Acknowledgement ………………………………………………………. ii
Abstract …………………………………………………………………. iii
Chapter 1: Introduction .............................................................................. 1
Background of the Study ........................................................... 2 Significance of the Study ........................................................... 3
Statement of the Problem ........................................................... 3 Scope and Delimitation ............................................................... 4
Chapter II: Review of Related Literature and Study
Review of Related Literature ....................................................... 5-6 Review of Related Study .............................................................. 7-9 Conceptual Framework ................................................................. 10 Null Hypothesis ............................................................................. 11 Definition of Terms ...................................................................... 11-12
Chapter III: Methodology
Materials ........................................................................................ 13
Instrument Used ............................................................................ 13 Procedure ....................................................................................... 13-14 Data Gathering Procedure ............................................................... 15


Page
Chapter IV: Results and Discussion
Tables 1.1 – 1.4 ..................................................................................... 16-18
Statistical Test ....................................................................................... 19-21
Chapter V:
Summary ................................................................................................. 22
Conclusion ............................................................................................... 22-23
Recommendation .................................................................................... 23
Appendix .............................................................................................................. 24-28
Bibliography ....................................................................................................... 29

Chapter I
Introduction

Acacia is a genus of shrubs and trees belonging to the subfamily Mimosoideae of the family Fabaceae, first described in Africa by the Swedish botanist Carolus Linnaeus in 1773.
Acacias are also known as thorn trees or wattles, including the yellow-fever acacia and umbrella acacias.
There are roughly 1300 species of Acacia worldwide, about 960 of them native to Australia, with the remainder spread around the tropical to warm-temperate regions of both hemisphere, including Africa, Southern Asia and the Americas. (http://en.wikipedia.org/wiki/Acacia_tree)
Dye can be generally be described as a colored substance that has an affinity to the substrate to which it is being applied. The dye is generally applied in an aqueous solution, and may require a mordant to improve the fastness of the dye on the fiber.
(http://en.wikipedia.org/wiki/Dye)
1
Background of the Study
The Acacia Bark is usually in curved pieces, externally grayish brown, darkening with age, often with irregular longitudinal ridges and sometimes transverse cracks. Its inner surface longitudinally striated, fracture irregular and coarsely fibrous. It also has a slight tan-like odor and astringent taste.
Acacia’s bark contains from 24 to 42 percent of tannin and also Gallic acid. Because of these constituents the researcher are inspired to produce a dye from it. Medicinally it has special use in diarrhea, mainly in the form of a decoction, the British Pharmacopoeia preparation being 6 parts in 100 administered in doses of ½ to 2 fluid ounces. The decoction also used as an astringent gargle, lotion or injection.
(http://www.botanical.com/botanical/mgmh/a/acaci003.html)
Dyeing, process of coloring materials, such as textile fibers, so that the coloring matter becomes an integral part of the fiber. Dyes, or dyestuffs, are soluble compounds that can be either absorbed and retained by the fiber or chemically combined with it. Dyes are generally fast—that is, they retain their color in the fiber throughout the textile-making process and under exposure to normal wear, including sunlight, water, and detergent washing.
(Microsoft ® Encarta ® Premium Suite 2005. © 1993-2004 Microsoft Corporation. )
As what was everyone observed, majority of the people today are using dye which made by other people, specifically the Chinese, which means that our local enterprising will undergo competition. That is why, as one of the responsible citizens in this country, the researchers are determined to make a high quality dye not only to sustain the needs of every Filipinos but also to help the economy of this country to stabilize through marketing this dye.
2
Significance of the Study

This research study is significant for another newly-discovered alternative dye using the bark of an acacia tree. As a possible result, dye prices in markets are expected to decline due to its uprising discoveries. The formulation of unique or probably improved research investigations concerning such matter together with the advancement of technology which is greatly influenced by the rapid westernization of the country could contribute mainly upon the solution of the severe illness the country is currently facing which is the undeniable poverty.
This could be an effective fabric coloring which would surely promote durability and long-lasting efficacy and satisfaction.

Statement of the Problem
This research study aims to answer the following questions:
1.) What is the color of the dye from the Acacia bark?
2.) Is there any significant difference between the long-lasting effect of acacia bark as dye applied on the fabrics with mordant and on fabrics without mordant in terms of color of the dye?
3.) Is there any significant difference between the effect of the color on the fabric of acacia bark as dye on the fabrics with and without mordant?

3

Scope and Limitation
The scope of this study is to determine the effectiveness of acacia bark used as dye on different types of fabrics with and without mordant and it is limited only in producing dye from acacia bark.
This study was conducted at Iligan City National High School by the researchers namely Carmille Soliel C. Tabanao, Darlyn E Tabañag and Catherine Grace N. Anggo Fourth Year Students, Section Einstein of Science Curriculum S.Y. 2007-2008.

4
Chapter II
Review of Related Literature and Review of Related Study

Review of Related Literature
Dyeing, process of coloring textile fibers and other materials so that the coloring matter becomes an integral part of the material rather than a surface coating. Dyeing consists of immersing the textile or yarn in a solution of a dyestuff and raising the solution temperature to enable sufficient dyestuff to pass from the solution into the fibers. The textile material and the dye solution selected must have an affinity. If a consistent color is to be produced, the dye liquid must be evenly applied, and must permeate the material thoroughly. The material must therefore be moved continually, as the even wetting of the material by the dye liquid is crucially important. Printing is a process allied to dyeing in which a fixed pattern is applied to one side of the fabric by means of a dyestuff carrier paste.
Dyeing is a complex, specialized science. Today nearly all dyestuffs are synthetically produced compounds, and in most instances surpass natural dyes.
(Microsoft ® Encarta ® Premium Suite 2005. © 1993-2004 Microsoft Corporation. )
Acacia, important genus of trees or shrubs of the legume family. Most of the 1,200 species of the genus are native to tropical regions and Australia. The normal type of leaf is bipinnate (divided into leaflets), but it is often modified, especially among the species that have had to adapt to the intense heat and drought of Australia. Some Australian species have modified leaves called phyllodes.
5
Many species have hooks or thorns, the latter are sometimes enlarged at the base and house ants that deter other climbing plants and other insects. The genus is of great and varied economic importance, yielding edible seeds, and valuable timber and gum.
(Microsoft ® Encarta ® Premium Suite 2005. © 1993-2004 Microsoft Corporation. )
Acacia bark is a usually curved piece, externally grayish brown, darkening with age, often with irregular longitudinal ridges and sometimes transverse cracks. Inner surface longitudinally striated, fracture irregular and coarsely fibrous. It has a light tan-like odor and astringent taste. (http://www.botanical.com/botanical/mgmh/a/acaci003.html)
The bark is hard and woody, rusty brown and tending to divide into several layers. The outer surface of colder pieces is covered with thick blackish periderm, rugged and fissured. The inner surface is red, longitudinally striated and fibrous. (http://www.botanical.com/botanical/mgmh/a/acaci003.html)
It contains from 24 to 42 percent of tannin and also Gallic acid. Its powerful astringency causes it to be extensively employed in tanning. (http://www.botanical.com/botanical/mgmh/a/acaci003.html)

6
Review of Related Study
Walnut as Dye
Description
Black walnut trees can grow from 45 to 75 feet tall. They are deciduous trees whose leaves turn a bright yellow in the fall. Older trees have bark which has broad, irregular ridges. Black walnut trees can often be found growing along creek beds.
Black walnut leaves are divided into 9 to 19 leaflets which are lance-shaped. Nestled in among the leaves you can find the walnuts, enclosed in their green husks.
INGREDIENTS
*2 1/2 gallons whole walnuts inside their green husks
12 oz. mordanted wool
12 quarts water
METHOD
1) Cover walnuts (with their husks on) with water, and soak 3 days.
2) Boil 2 hours.
3) Cool a few hours. Add water if needed so yarn won't be crowded.
4) Add presoaked, mordanted yarn. Boil 2 hours.
5) Cool in dye bath.
6) Rinse.
7) Dry in shade.
(http://home.onemain.com/~crowland/Pages/Walnut.html)
7
Beets as Red Dye
Materials
• 2 pots
• Beets
• Wool or fleece
• Alumn
• Strainer

Steps
1.) Turn on your stove burner to high. Fill a pot with about 4 quarts of water and place it on the stove. Allow the water to come to a full boil.
2.) Clean and cut up about 2 pounds of beets. Cut them into small pieces.
3.) Place the beets into the boiling water carefully and allow to boil for an hour. Watch the water turn red as the water boils.

4.) While your waiting for the beet dye to finish boiling, prepare the fabric that you wish to dye (wool or fleece is best) so the fabric will retain the color. In a stainless steel pot, dissolve 5 ½ oz of alum (aluminum potassium sulfate) in 4 gallons of warm water.

8
5.) Place your wet fabric into the alumn mordant mixture and stir with a wooden spoon. Boil it in the mixture for 20 minutes. Remove, cool and wring out the fabric.

6.) Using a strainer, strain the beets from the dye mixture after it has boiled for an hour.
Throw out the beets and keep the red dye mixture simmering on the stove.

7.) Place the wet fabric from the alumn mordant mixture into the dye. Boil the fabric in the dye for 1 hour or until it turns red.
(http://www.ehow.com/how_2079352_make-red-dye-from-beets.html)

9
Conceptual Framework:

Dye from Acacia bark Extract
on Katrina Fabric with
mordant
 effect of the color on
Experimental Set-up fabric
it’s long-lasting effect
color of the dye

Dye from Acacia bark Extract
On Katrina Fabric without
mordant ExperimentalSetup
10
Null Hypothesis

1) There is no significant difference between the long-lasting effect of acacia bark as dye applied on the fabrics with mordant and on fabrics without mordant in terms of color of the dye.
2) There is no significant difference between the effect of the color on the fabric of acacia bark as dye on the fabrics with and without mordant.

Definition of Terms
1.) Affinity  a measure of the likelihood of a chemical reaction taking place between two substances.
2.) Bark  hard covering or rind of the stem, branches, and roots of a tree or other plant, as distinguished from the wood.
3.) Bipinnate  used to describe leaves divided into leaflets that are themselves subdivided
4.) Dyes  are soluble compounds that can be either absorbed and retained by the fiber or chemically combined with it: can be generally being described as a colored substance that has an affinity to the substrate to which it is being applied.
5.) Fabric  cloth of any type made from thread or fibers, whether woven, knitted, or felted.
6.) Gallic Acid  a colorless crystalline solid that occurs in plants and is obtained from tannin. It is used as a tanning agent, in making inks and paper, and in photography.

11
7.) Long-lasting effect  the number of times the Katrina fabric was washed after dyeing process.
8.) Mordant  a substance that fixes a dye in and on textiles and leather by combining with the dye to form a stable insoluble compound.
9.) Phyllode  a flat leaf stalk that functions as a leaf in certain plants, such as the acacia.
10.) Tannin  a brownish or yellowish substance found in plants and used in tanning, dyeing, and as an astringent.
12
Chapter III
Methodology

Materials:

 2 pieces of 200 mL Beakers
 Tripod
 Wire Gauze
 Alcohol Lamp
 Weighing scale
 70% solution of Alcohol
 2 Bottles of Denatured Alcohol
 4 pcs. of Katrina cloth 4 x 4 inches
 Ferrous Sulfate
 Acacia Bark
(Outer part of the bark)

Procedure:
Gathering of Materials:

Prepare the needed materials and the apparatuses. Be sure that the materials are thoroughly cleaned, including the dried acacia bark and the Katrina cloth. Katrina should have four pieces with the sizes of 4 x 4 inches. Separate the four pieces of Katrina fabric so that you will have four sets. Let set 1 and set two be applied with mordant but set two will be washed with detergent to determine the long lasting effect while set three and set four without mordant but set four will be washed with detergent same as set two.

13
A. Extracting Dye from Acacia Bark:
Measure the volume of alcohol which is about 175 mL. and weigh 100 grams of acacia bark. Soak the acacia bark in the beaker with alcohol. Cover the beaker and leave it for about 2 hours. After soaking, boil it with the use of an alcohol lamp until you can get the extract. Let it cool.

B. Applying Mordant:
Get a Petri dish and put water unto it. Mix the Ferrous Sulfate to produce a mordant. Then, apply an appropriate amount of mordant on the fabrics you selected to be applied with mordant.

C. Evaluating the efficacy of Acacia bark as Dye:
First, soak the fabric without mordant into the cooled acacia bark extract. Leave it for 15 minutes. After soaking the fabric, remove it from the beaker. Hang to dry. Then, soak the other fabric without mordant into the extract and leave it for 15 minutes. Let it dry. After drying, wash it with detergent and let it dry. Repeat the process for at least two days. Next, soak the other fabric with mordant into the cooled acacia bark extract. Leave it for 15 minutes. After soaking the fabric, remove it from the beaker. Hang to dry. Then soak the fabric with mordant. After drying, wash it with detergent and dry it again. Repeat the process for at least two days. This is done to test the long-lasting effect of the dye from the acacia bark extract. Get the comparison of the Fabrics with and without mordant through a survey. Determine which of the four pieces does acacia bark’s dye will show the best result on both set-ups – with and without mordant.

14
Data Gathering Procedure
Survey Questionnaire - 30 respondents will be ask to answer the question about the color of the dye, effect of the color on the fabric and the long-lasting effect of dye on the fabric.

15
Chapter IV
Results and Discussion

This Table shows the number of respondents who gave their comment regarding the color and long-lasting effect of acacia bark as dye applied on the Katrina cloth with mordant.
Table 1.1
30 Female Respondents
Katrina (with mordant)
Color Long-lasting Effect
01  
02  
03  
04  
05  
06  
07  
08  
09  
10  
11  
12  
13  
14  
15  
16  
17  
18  
19  
20  
21  
22  
23  
24  
25  
26  
27  
28  
29  
30  

16
This Table shows the number of respondents who gave their comment regarding the color and long-lasting effect of acacia bark as dye applied on the Katrina cloth without mordant.
Table 1.2
30 Female Respondents Katrina (without mordant)
Color Long-lasting effect
01  
02  
03  
04  
05  
06  
07  
08  
09  
10  
11  
12  
13  
14  
15  
16  
17  
18  
19  
20  
21  
22  
23  
24  
25  
26  
27  
28  
29  
30  

Legend:

  Excellent   Very Good

  Undecided   Poor

17

These two tables shows the comments of the 30 female respondents in percentage form regarding the color and long-lasting effect of the acacia bark’s dye applied on the Katrina fabric with and without mordant.
With Mordant Without Mordant
Table 1.3 Table 1.4
30 Female

Respondents Katrina

Color

Excellent (  ) 33.33%

Very Good ( ) 63.33%

Undecided (  ) 3.33%

Poor (  ) 0%

Long-lasting Effect

Excellent (  ) 13.33%

Very Good ( ) 13.33%

Undecided (  ) 46.67%

Poor (  ) 26.67%
30 Female

Respondents Katrina


Color

Excellent (  ) 23.33%

Very Good ( ) 73.33%

Undecided (  ) 3.33%

Poor (  ) 0%

Long-lasting Effect

Excellent (  ) 23.33%

Very Good ( ) 60%

Undecided (  ) 16.67%

Poor (  ) 0%


This table implies that the acacia bark’s dye is more effective on the Katrina cloth without mordant in terms of color with a percentage of 33.33%
This table also shows that the acacia bark’s dye is more effective on the Katrina cloth with mordant in terms of long-lasting effect with a percentage of 23.33%

18
The Statistical Calculation for Quality Test

This table shows the Statistical Calculation for the Quality Test on the effects of Acacia bark’s Dye on the Katrina cloth with and without mordant in terms of it’s color and long-lasting effect.

Table 1.5
Effects of dye Katrina Cloth Total

Katrina fabric with mordant Color Long-lasting Effect 6.27
3.2 3.07
Katrina fabric without
mordant 3.3 2.13 5.43

Total
6.5 5.2 11.7


a. Ho: There is significant difference on the quality test of the fabrics with and without mordant (Katrina).

b. H1: There is no significant difference on the quality test of the fabrics with and without mordant (Katrina).

c. X2 > 7.815 ; Mean = 30

d. Use 0.05 level of significance

19
e. Computation:

For 3.2: (6.5)(6.27) = 3.483
11.7

For 3.07: (5.2)(6.27) = 2.7867
11.7

For 3.3: (6.5)(5.43) = 3.0167
11.7

For 2.13: (5.2)(5.43) = 2.413
11.7

Table 1.1 Results using Chi- Square Distribution

OF EF OF - EF ( OF-EF ) 2 ( OF – EF ) 2/EF
3.2 3.483 -0.283 0.08 0.023
3.07 2.7837 0.2833 0.08 0.029
3.3 3.0167 0.2833 0.08 0.027
2.13 2.413 -0.283 0.08 0.033
∑ ( OF – EF ) 2/EF = 0.112

f. Decision:

Ho – accepted : since / 0.112 / is less than / 7.815 /. Therefore there is significant.

H1 - rejected : difference on the quality test of the fabrics with and without mordant
(Katrina)
20

STATISTICAL TABLE
TEST MEAN Level of Significance Computed Value Critical Value DECISION
Chi-square 30 0.05 0.112 7.815 Ho=accepted
H1=rejected

This Statistical table shows the use of Chi-square to test the significance of the datas. The null hypothesis states that there is a significant difference on the color and long-lasting effect on the acacia bark extract applied on the Katrina fabric.

21
Chapter V
Summary, Conclusion and Recommendation

SUMMARY
This study was conducted at Iligan City National High School; ESEP Building last February 23, 2006 and was conducted to produce a dye from the Acacia Bark and its efficacy as a black dye for cloth. This was produced through extraction then the Katrina fabrics were submerged for testing.
After conducting the experiment, the researchers made an evaluation regarding the result through a sensory evaluation or survey.

CONCLUSION
Based on the result of the Chi-square Test, there is a significant difference on the quality test of the fabrics with and without mordant (Katrina cloth) since the value of
∑ (OF-EF) 2/EF is equal to 0.122 and it is within the range which is x2>7.815.
Based on the conducted experiment, the dye from the Acacia bark is black-colored.
According to the sensory evaluation conducted by the researchers, there is a significant difference on the long-lasting effect of Acacia bark extract as dye applied on the Katrina cloth with mordant and on the Katrina cloth without mordant. There is a significant difference on the effect of the color of Acacia bark’s dye on the fabrics with and without mordant.
22
Therefore, the Acacia bark can be a good source of dye. Through a survey, the researchers determined how effective the Acacia bark as dye is.


RECOMMENDATION
The researchers would like to recommend that:
1) The acacia bark extract can also be used in making an ink.































23
The Researchers
















Name: Tabanao, Carmille Soliel Casuco
Surname First Name Middle Name

Nickname: Cam2x and Carm Sex: Female

Birth date: February 13, 1991

Age: 17 yrs. Old

Religion: Roman Catholic

Address: Purok 1 Merila, Iligan City

Cell # or Tel. # : 0927691087




26



















Name: Anggo, Catherine Grace Nillas
Surname First Name Middle Name

Nickname: Grace Sex: Female

Birth date: February 29, 1992

Age: 16 yrs. Old

Religion: Latter Day Saints

Address: Zone 5 Del Carmen, Iligan City

Cell # or Tel. # : 09164339801


27




















Name: Tabañag, Darlyn Entrampas
Surname First Name Middle Name

Nickname: Ate Darl Sex: Female

Birth date: March 16, 1991

Age: 16 yrs. Old

Religion: Jehovah’s Witnesses

Address: Purok 5 Zone 6 Fuentes Maria Cristina, Iligan City

Cell # or Tel. # : 09168263854

28
Bibliography



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“Acacia Tree”.(Microsoft ® Encarta ® Premium Suite 2005. © 1993-2004 Microsoft Corporation. )
“Acacia bark “.(http://www.botanical.com/botanical/mgmh/a/acaci003.html)
“Dye”.(http://en.wikipedia.org/wiki/Dye)
“Dye”.(Microsoft ® Encarta ® Premium Suite 2005. © 1993-2004 Microsoft Corporation. )
“Dye”.(http://home.onemain.com/~crowland/Pages/Walnut.html)
“Dye”.(http://www.ehow.com/how_2079352_make-red-dye-from-beets.html)