Annatto (Bixa orellana L) Ink

Research I-B Adviser: Mrs. Ramonita F. Calumpang

Researcher:

Marcelle Antonette A. Carriaga

 Abstract This study was launched to produce a natural ink. The chief ingredients of the said product is Annatto(Bixa orellana L) seeds, the researcher decided to name as Annatto Ink. Three set-ups were organized by the researchers and each set-up had three trials. The researchers gathered all the equipments and ingredients required, namely: Annatto seeds, vinegar, rolling pin/knife, strainer, container, kettle/sardine can, pliers(if using sardine can), chopping board. The procedure the researchers conducted was simple. First, all the ingredients were prepared. Next, soak the annatto seeds. After soaking, pulverize the soaked annatto seeds. After, boil the annatto seeds with the vinegar. After boiling, pour it in a container. Ten respondents were requested to judge and rate the product in terms of color, odor, and appearance. The respondents then rated each trial from the scale of one (1) as the lowest and ten (10) as the highest rating. After the statistical analysis was computed, the researchers concluded that the best set-up is Set-up C, trial 3. The researchers then concluded that in making ink, one could also use annatto seeds. Set-up C (trial 3) being the best set-up proved to be the having the good quality of color, having good odor, and most appealing among the other set-ups.

 Acknowledgement Upon making this investigatory project, I the researcher had encountered many problems. In the midst of all these difficulties; I still made this a successful one. But this would not be possible without the endless or never ending support of some people whom I consider now my inspiration. First and foremost, I would like to give it all back to our Heavenly Father, thank You for bestowing the researchers the said essential individuals that made this study successful and for giving the researchers a precious gift and this is their determination to give their best to make their study triumphant. All the glory belongs to You! I would also express my profound thanks and gratitude to my dear parents, Mr. Venerando C. Carriaga and Mrs. Jo-ann Carriaga. I acknowledge them for their words of encouragement, for their candid, sharp and positive criticism and for the financial support throughout the whole duration of my investigatory project. To Mrs. Ramonita F. Calumpang, our Research I-B adviser, thank you for the approval of my study and for your additional suggestions in our product.

 Table of Contents

Title Page . Abstract ... Acknowledgement ... Introduction ................................................................................................................. A. Background of the Study .. B. Statement of the Problem/ Objectives . . C. Significance of the Study .. D. Scope and Limitations .. E. Review of Related Literature F. Related Studies .. Methodology .................................................................................................................

Results and Discussion .................................................................................................

Conclusion ....................................................................................................................

Recommendations .........................................................................................................

Bibliography ..................................................................................................................

I.

INTRODUCTION

A. Background of the Study Nowadays our country has big problem about our economy. Everyone to solve it, but what do you think are the possible ways to prevent his crisis? I suggest that being practical is one of the solution to this problem. As students, we can be practical by using our knowledge instead of wasting our money. This project do not need much time and money because we can make our own ink within a short period of time. The basic ingredients are vinegar and annatto seeds.The seeds of the Annatto are used locally for coloring food. The coloring matter of the fruit, Annatto is employed commercially for coloring butter and in preparation of various polishers for russet leather. B. Statement of Problem/ Objectives 1. General Objective The study aims to produce an ink. 2. Specific Objective This study aims to produce an ink using achiote seeds. C. Significance of the Study The importance of the study can be observed on how to produce ink out of achiote seeds. By this, it may lessen the expenses of buying ink.

D. Scope and Limitations The production of this ink depends on the availability of the achiote seeds. The color produced by this ink is only color Red.

E. Review of Related Literature Annatto/Achiote Annatto, sometimes called roucou or achiote, is a derivative of the achiote trees of tropical regions of the Americas, used to produce a yellow to orange food coloring and also as a flavoring. Its scent is described as "slightly peppery with a hint of nutmeg" and flavor as "slightly nutty, sweet and peppery". Annatto coloring is produced from the reddish pericarp or pulp which surrounds the seed of the achiote (Bixa orellana L.). It is used as coloring in many cheeses (e.g., Cheddar, Gloucester cheese, Red Leicester, Gouda and Brie), margarine, butter, rice, custard powder, ice-cream, and smoked fish. Although it is a natural food colorant, it has been linked to cases of foodrelated allergies. Annatto is commonly found in Latin America and Caribbean cuisines as both a coloring agent and for flavoring. Central and South American natives use the seeds to make a body paint and lipstick. For this reason, the achiote is sometimes called the "lipstick-tree". Achiote originated in South America and has spread in popularity to many parts of Asia. It is also grown in other tropical or subtropical regions of the world, including Central America, Africa and Asia. The heart-shaped fruit are brown or reddish brown at maturity, and are covered with short, stiff hairs. When fully mature, the fruit split open, exposing the numerous dark red seeds. While the fruit itself is not edible, the orange-red pulp that covers the seed is used to produce a yellow to orange commercial food coloring. Achiote dye is prepared by grinding seeds or simmering the seeds in water or oil.

History and Mythology of the Annatto/Achiote The Latin name of this plant 'Bixa orellana' does not give much of a clue regarding its properties. The genus name is probably derived from the Portuguese 'biche' meaning beak which alludes to the beak shaped seedpods, while the species name is given in memory of Francisco de Orellano, a Spanish conquistador of the 16th century, who accidentally discovered the Amazon. Although the fruit of the Annatto tree are inedible it is often cultivated for its flowers and more especially for its seedpods. The pulp of the Annatto fruit yields a bright red dye, which has long been used both as a body paint and dye stuff for textiles or food. The ancient Maya and Aztecs regarded it as a symbolic substitute for blood and thus ascribed to it sacred connotations. It was also used to make ink and virtually all the ancient Maya scriptures were penned in annatto juice. The seeds also have a reputation as a female aphrodisiac and are believed to make bulls used for bullfighting more aggressive. The whole tree has a long history as a valued medicinal plant that has been used to treat a wide variety of conditions from fevers to cancer. Indigenous people still use the pulp for 'cosmetic purposes', as hair dye or lip stick, hence the English common name 'Lipstick tree'. The pulp is also said to repel insects and to protect against sunburn due to the UV-filtering properties of the carotenoid pigment known as Bixin. Its use as a food dye is just as ancient. The Aztecs were known to add Annatto to their sacred xocolatl brew and other foods. Its use as a food dye has persisted until today. Annatto is probably one of the most ubiquitous of all food dyes used by the food industry. It lends its reddish tint to cheeses, butter and spreads, candy and custards. It is

also still used as a traditional food dye for meats. This use is most prevalent in the Philippines and in Central America and Mexico. (The bright red colour of Chinese poultry however is due to treatment with a caramelised malt solution.) The seed pods are processed by separating the pulp form the seeds, which are washed and used separately as a mild spice. A spice paste known as 'Achiote Recado' is a popular flavouring in Yucatan cuisine (southern Mexico). The meat is marinated in the paste and wrapped in banana leaves. Fish, chicken and especially pork or suckling pig can be treated this way. Even though Annatto is one of the most widely used food colouring substances of the food industry, some people appear to be highly allergic to it and lobby against the use of this additive. The way in which commercial annatto is processed as a dye involves hexane extraction, which just may possibly have something to do with these reported allergic reactions. Furthermore, the colouring agent, known as Bixin can now be produced by bio-engineering. Scientists have figured out the biochemical pathway and manipulated E.coli bacteria to produce Bixin. It might be interesting to conduct a comparative study of allergic reactions between, a) completely naturally processed annatto (see recado recipe below), bio-engineered bixin or commercially extracted annatto dye. Annatto dye is also used to colour hair-oils, shoe polishes, floor polishes, nailgloss, furniture, brass-lacquer, soap, cosmetics and pharmaceutical ointments as well as textiles, wool, leather and calico. The spiny, lipstick-red fruits were used by the indigenous peoples of Central and South America to make body paint, a pigment for mural paintings, and an ink. It was also used to

colour butter and cheese. In Belize, annatto is used to colour the rice red. Amazon tribes use it for body paint and as a protection from insects. They also use it as an aphrodisiac.

Annatto/Achiote Uses Annatto has had many uses over the centuries in Latin America and the Caribbean, including as a food dye, body paint, treatment for heartburn and stomach distress, sunscreen and insect repellent. In the Philippines, it is called atsuete and is used as food coloring in traditional dishes. Many Latin American cuisines traditionally use annatto in recipes of Spanish origin that originally call for saffron; for example, in arroz con pollo, to give the rice a yellow color. In Venezuela, annatto (called locally onoto) is used in the preparation of hallacas, perico, and other traditional dishes. In Brazil, both annatto (the product) and the tree (Bixa orellana L.) are called urucum, and the product itself may also be calledcolorau. Annatto/Achiote as Food Coloring As a food additive, annatto has the E number E160b. The fat soluble part of the crude extract is called bixin, the water soluble part is callednorbixin, and both share the same E number as annatto. Annatto seed contains 4.5-5.5% pigments, which consists of 70-80% bixin. In the United States, annatto extract is listed as a color additive "exempt from certification" and is informally considered to be a natural color. The yellowish orange color is produced by the chemical compounds bixin and norbixin, which are classified as carotenoids. However, unlike beta-carotene, another well-known carotenoid, they do not have the correct chemical structures to be vitamin A precursors. The more norbixin in

an annatto color, the more yellow it is; a higher level of bixin gives it a more reddish shade. Unless an acid-proof version is used, it takes on a pink shade at low pH. Cheddar cheese is often colored, and even as early as 1860, the real reason for this was unclear: English cheesemaker Joseph Harding stated "to the cheese consumers of London who prefer an adulterated food to that which is pure I have to announce an improvement in the annatto with which they compel the cheesemakers to colour the cheese". One theory is that cheeses regarded as superior in the 16th century had somewhat yellow color, possibly from high levels of carotene in the grass on which the dairy cattle fed. Producers of inferior cheese added annatto to the milk to make the cheese appear better quality, thus to command a higher price. As a food color, annatto has fewer tendencies to oxidize than beta carotene. Solvent-extracted annatto pigment present in edible oils at even low practical use levels, markedly delays polymerization of the oils during heating, and thus delays the development of the unhealthy by-products of polymerization. Whether this effect is also present in oil-extracted annatto pigment, where annatto seeds are held in edible oil at high temperature under near vacuum or inert gas, a process that may itself induce polymerization, is not known. Annatto appears to be one of the richest natural sources of delta-tocotrienol (a type of vitamin E), and contains as well gamma-tocotrienol (the ratio between gamma and delta forms is around 9-to-1), while not containing any tocopherols, which inhibit metabolism of tocotrienols in humans. Annatto is used as a source of these two types of tocotrienols for dietary supplements.

F. Review of Related Studies

Antimicrobial / Antifungal: (1) The study screened plants extracts - including Bixa orellana for antimicrobial activity and MIC. B orellana showed low MIC against against E coli and a better MIC against B cereus. Results showed that Bixa orellana, Justicia secunda and Piper pulchrum could be potential sources of new antimicrobial agents. (2) Study of organic extract of BO showed to possess a narrow spectrum of antimicrobial activity, effective only against Gram+ bacteria used in the study. Antibacterial / Antioxidant / Antidiarrheal / Neuropharmacolic / Anticonvulsant/ Gastrointestinal Motility Effect: Preliminary pharmacologic studies on the methanol extracts of BO leaves showed statistical decrease in locomotor activity, anticonvulsant effect, analgesic and antidiarrheal effect and a delay i gastrointestinal motility. It also showed radical scavenging properties and antibacterial activity against agents of diarrhea and dysentery. Hypoglycemic / Antidiabetic: (1) Study of BO oil seed suspension of the red seed coat showed it to be a potent hypoglycemic agent. Extract study showed hypoglycemia in both normal and streptozotocin diabetic dogs. (2) Annatto extract was found to decrease blood glucose in fasting normoglycemic and streptozocin-induced diabetic dogs. The activity was attributed to the stimulation of peripheral utilization of glucose. Milk-based pigment: Study showed B orellana seeds can be used as a pigment in making milk-based paint. Anti-Gonorrheal: In a study of 46 plants popularly used in Guatemala for the treatment of gonorrhea, B orellana bark was one of the most active plants that inhibited strains of freshly isolated N gonorrhea.

 Antibacterial / Anti-Staphylococcal Activity: Study showed crude ethanolic extracts from leaves of B orellana and bark of Alstonia macrophylla showed potential antibacterial effect against S aureus. Aldose Reductase Inhibition: Study of hot water extracts of B orellana, C morifolim and I batatas showed potent inhibitory activity towards lens aldose reductase. Phytochemicals yielded isoscutellarein from B orellana as its potent inhibitor. y Atsuete Seeds, Alugbati Fruit, and Luyang Dilaw as Substitute for Typewriter Ribbon Ink Researchers: Don Carlo Inato, John Kris Magbanua, Kemuel Ed Kevin Madelo, Diamondrav Villamor, Aprilson Maroo Arnado, King Lustre Abstract The researcher aimed to develop an economically useful substitute for the commercially available typewriter ribbon ink. The project intends to determine the achievability of producing a typewriter ink from Atsuete seeds, Alugbati Fruits and Luyang Dilaw Extracts similar to the commercialized typewriterribbon ink in terms of clarity, cleanliness, general acceptability, glossiness, and goodness of the printed material. y The Feasibility of Atsuete as Ink Pigment

Researchers: Carlo Ace Cariaga, Marco Fernando, Darryl Aaron

II.

Methodology A. Materials/Equipment y y y y y y y y Annatto Seeds Vinegar Rolling pin/knife Strainer Container Kettle/Sardine can Pliers(if using a sardine can) Chopping board

B. Treatment/General Procedure The procedure the researchers conducted was simple. First, all the ingredients were prepared. Next, soak the annatto seeds. After soaking, pulverize the soaked annatto seeds. After, boil the annatto seeds with the vinegar. After boiling, pour it in a container. Set-up A Trials Annatto (grams) 1 2 3 100 200 300 Vinegar (mL) 50 50 50 No. of minutes soaked (minutes) 2 2 2

In set-up A, the amount of vinegar and the number of minutes soaked was the controlled variable. The manipulated variable in this set-up is the annatto seeds. The best trial in this set-up is set-up A trial 1. Set-up B Trials Annatto (grams) 1 2 3 100 100 100 Vinegar (mL) 100 150 200 No. of minutes soaked (minutes) 5 5 5

In set-up B, the amount of annatto seeds and the number of minutes soaked was the controlled variable. The manipulated variable in this set-up is the amount of vinegar. The best trial in this set-up is set-up B trial 1. Set-up C Trials Annatto (grams) 1 2 3 100 100 100 Vinegar (mL) 55 55 55 No. of minutes soaked (minutes) 2 5 10

In set-up C, the amount of annatto seeds and the amount of vinegar was the controlled variable. The manipulated variable in this set-up is the number of minutes soaked. The best trial in this set-up is set-up B trial 1.

III. RESULTS AND DISCUSSION A. Findings In set-up A, the best trial is trial 1, for set-up B is trial 1, and for set-up C is trial 3. To be able to know which among the set-ups is the best; the researchers let ten (10) respondents judge the products. Legend: 1-4 Needs Improvement Set-up A (Trial 1) Respondents 1 2 3 4 5 6 7 8 9 10 Total: Color 8 10 7 8 9 9 10 8 9 9 87 Odor 8 9 8 10 8 7 9 9 8 8 84 Appearance 9 7 9 8 8 10 9 9 8 9 86 5-8 Satisfactory 9-10 Excellent

Set-up B (Trial 1)

Respondents

Color

Appearance Odor

Appearance

1 2 3 4 5 6 7 8 9 10 Total:

8 7 10 9 10 9 8 10 8 9 88

8 9 9 10 7 8 7 9 8 9 84

9 7 8 8 9 8 8 8 8 9 82

Set-up C (Trial 3) Respondents 1 2 3 4 5 6 7 8 9 10 Total: Color 8 9 8 10 7 8 8 10 8 9 85 Odor 9 10 10 9 8 9 9 10 8 10 92 Appearance 8 8 7 9 8 9 8 10 8 8 83

Over-all Rating Set-up A B C TOTAL Color 87 88 85 260 Odor 84 84 92 260 Appearance 86 82 83 251 TOTAL 257 254 260 771

After letting the respondents rate the best trials of each set-up, set-up A (trial 2) got the highest average which clearly implies that it is the best among the other set-ups.

III.

CONCLUSION The researchers therefore conclude that in making ink, one could also use annatto seeds. Set-up C (trial 3) being the best set-up proved to be the having the good quality of color, having good odor, and most appealing among the other set-ups.

IV.

Reccomendations The researchers would like to recommend to the future researchers to think of ways on how to make the odor of the ink better and. It is also recommended to find other seeds as the main ingredient in making inks.

VI. BIBLIOGRAPHY http://scinet.dost.gov.ph/union/ShowSearchResult.php?s=2&f=&p=&x=&page=&sid=1&id=The +feasibility+of+atsuete+as+ink+pigment&Mtype=PROJECTS http://spaconline.org/academy/?page_id=51 http://www.stuartxchange.org/Asuete.html http://en.wikipedia.org/wiki/Annatto http://www.sacredearth.com/ethnobotany/plantprofiles/achiote.php http://www.cloverleaffarmherbs.com/annatto/