World Journal of Dairy & Food Sciences 6 (1): 71-78, 2011ISSN 1817-308X© IDOSI Publications, 2011

Corresponding Author: Ali Aberoumand, Department of Fisheries, Behbahan High Educational Complex, Behbahan, Iran.Postal address: No 22, second line of Ab va Barg, Zolfeghari Street, Behbahan, Khuzestan Province, Iran.Tel: +98-9166716540, Fax: +98-6712229969, E-mail: aaberoomand@yahoo.com.

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A Review Article on Edible Pigments Properties and Sources asNatural Biocolorants in Foodstuff and Food Industry

Ali Aberoumand

Department of Fisheries, Behbahan High Educational Complex, Behbahan, Iran

Abstract: Consumers are avoiding foods containing synthetic colourants, which lead food industries toreplace them by natural pigments. Food colorants may be classified into synthetic, nature-identical, inorganicand natural colorants. Natural colorants for food are made from renewable sources. Most often, the colorantsare extracted from plant material, but other sources such as insects, algae, cyanobacteria and fungi are usedas well. Natural colorants are usually extracted and concentrated using either water or lower alcohols forwater-soluble pigments and organic solvents for lipophilic pigments. Legislation restricts which colorantsare allowed, what sources may be used for that particular colorant, what solvents may be used to extract it andthe purity of the pigment. Colorants are formulated to make them more suitable for a variety of foods and drinksand to increase their stability.

Key words: Food colorants % Natural colourants % Pigments % Properties

INTRODUCTION Examples of inorganic colors are titanium dioxide, gold

Color is an important characteristic of food. Based on from a variety of sources, natural food colorants can bethe color of the food, first impressions are made: Is the grouped into a few classes, the three most important offruit immature, ripe, or overripe? Is the toast burnt? Is the which are: tetrapyrrols, tetraterpenoids and flavonoids.food fresh? Based on these first impressions, a judgment The most important member of the tetrapyrrols isis made whether the food is safe to eat or not and chlorophyll, which is found in all higher plants.whether it can be expected to taste good or not. Since Carotenoids are tetraterpenoids that are as ubiquitous ascolor is closely associated with expectations, the addition chlorophyll, since they too are part of the photosyntheticof color to food is a way to fulfill these expectations. apparatus. They also give the yellow–orange–red color ofColor is added to food for one or more of the following many fruits. Anthocyanins are a group of flavonoidsreasons: (1) to replace color lost during processing, (2) to that provide the red–purple shade of many fruits, inenhance color already present, (3) to minimize batch- particular berries (e.g., strawberries, elderberries andtobatch variations and (4) to color otherwise uncolored black currants). Other important classes of colorants arefood. Food colors can be divided into four categories: (1) the anthraquinones (carmine, lac, kermes and madder)natural colors, (2) nature-identical colors, (3) synthetic and the betalains (beetroot). The terms “pigment” andcolors and (4) inorganic colors. Natural colors are “dye” are often used interchangeably. Strictly speaking,pigments made by living organisms. Usually, pigments a pigment is insoluble in the given medium, whereas amade by modification of materials from living organisms, dye is soluble. Thus, carotenoids are dyes in oil butsuch as caramel, vegetable carbon and Cu-chlorophyllin pigments in water. This distinction may be difficult to(vide infra), are also considered natural though they are in maintain if nothing is assumed about the medium and infact (except for carbon) not found in nature. Nature- the following the term “pigment” will be used for coloredidentical colors are man-made pigments which are also substances in general [1-8].found in nature. Examples are carotene, canthaxanthin The development of foods with an attractiveand riboflavin. Synthetic colors are man-made colors appearance is an important goal in the food industry.which are not found in nature, these are often azo-dyes. Increasingly, food producers are turning to natural food

and silver. Although structurally very diversified and

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colors, since certain artificial color additives have this family plant displayed a broader color spectrum anddemonstrated negative health issues following their were devoid of the mention drawbacks in red beet [18].consumption [9]. Because of the deficiencies of existing Selected species of cactus fruits recognized asnatural food colorants, the demand for natural pigments purple-fleshed Hylocereus polyrhizus have very recentlyis repeatedly raised by the food industry. This demand been suggested as promising betacyanin source. Thecan be fuelled by research to offer a more natural-healthy primary findings on the pigment potential and the scarceway of coloring foods and provide a clean label knowledge about color characteristics encouraged moredeclaration. Therefore, part of plant pigment research is investigations. Preliminary data from different Hylocereusseeking new sources of pigments. This will not only genotype were promising and further investigations aredirected in finding natural alternatives for synthetic dyes, needed for process optimisation with respect to colorbut also discover new procedures for the pigment shade and pigment yield. The main obstacle in theseproduction. The diversity of tropical and subtropical fruits is the pectic substances. Method of degradationvegetation offers a promising range of unknown plant of pectic material is necessary to facilitate pigment release,compounds that might prove applicable for the needs of increasing yield and reducing processing wastes. Inhumans. The most common plant pigments are Malaysia fruits of Hylocereus polyrhizus (genuscarotenoids, chlorophylls, anthocyanins and betalains. Hylocereus, Cactaceae) have been cultivated as fruit cropMost research has been focused on carotenoids and locally. Red color pigments from fruits of H. Polyrhizusanthocyanins but betalains have recently gained interest could be an interesting possibility as alternative toin food science. Commonly broad arrays of anthocyanin betanin from red beet. In view of the fact that most naturalcontaining extracts are used for food coloring purposes; food colorants are supplied as concentrated juices orso far, there is only one single betalainic source that has extracts and to get a better understanding of how pigmentbeen extensively used in the food industry worldwide. profile and hue are related in making decision forCompared to anthocyanins, betalains are ideal for coloring commercial applicability, the present work aimed to:low-acid foodstuff as they maintain their color over a widearray of pH, from 3 to 7. The most important betalainic C To extract betacyanin pigments from pulp and skin ofsources for natural red coloring are selected varieties of Hylocereus polyrhizus.red beet, commercial preparations of which are mainly C To identify the constituents of the betacyanincomposed of the red-purple betanin and its C15-isomer pigments from pulp of Hylocereus polyrhizus.isobetanin. C To evaluate the effect of enzyme treatment on yield

Betacyanins are important constituents of betalain of betacyanin constituent.pigments. They are the main compounds associated with C To compare the color of extract from Hylocereusthe red color exhibited by flowers, fruits and other plant polyrhizus to that of red beet [19-21].tissues. Red beet has been established in the market asthe oldest and most abundant red food colorant, called Natural Distribution of Pigments: Chlorophylls andbetanin, that is known as E-162 in the European Union carotenoids are the most abundant pigments in nature.[10-13] and as 73.40 in the chapter 21 of the Code of They are involved in fundamental processes and life onFederal Regulations (CFR) section of the Food and Drug Earth depends on them. Plants, photosynthetic bacteriaAdministration (FDA) in the USA (Griffiths, 2005). and protozoa (plankton) are the main sources of thePractically it has been used to color foods such as organic materials that are required for the developmentyoghurt, confectionery, ice creams, syrups, sausages and of other living organisms such as vertebrate andprocessed meats. However, the typical earthy flavor invertebrate animals. Chlorophyll is not found in animalscaused by geosmin and high nitrate concentrations but carotenoids accumulate in some organs (e.g., eyes)associated with the formation of carcinogenic and tissues (e.g., skin of fish, bird plumage). In general,nitrosamines [14, 15] may affect its commercial use. animal carotenoids are obtained from the common diet.Furthermore, the risk to carry-over earth-bound Other pigments are also found in animals; some havemicroorganisms from the raw material in red beet is a important functions (e.g., heme proteins, riboflavin),crucial point [16,17]. Thereby, there is a high demand for whereas the function of others is not yet completely clearother alternative compounds that can be substitute for red (e.g., melanins, flavonoids). Other organisms havebeet. In recent years fruits from the Cactaceae have been interesting pigments that have been used or haveintroduced as a promising betalain source. The fruits of potential use. Lichens produce depsides, the ancient

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and most extensively used dyes which were used as is mentioned in the Bible [26]. Use of natural biocolarantstextile dyeing agents. In addition, they have application in food is known from Japan in the shosoin text of theas sunlight filters, as chemical indicators (litmus paper, Nara period (8th century) contains references to coloredpH indicators) and as cytological stains. Some of the soybean and adzuki-bean cakes, so it appears thatpigments obtained by treatment of lichen substances colored processed foods had been taken at least byare orcein and parietin: More than 1000 pigments people of some sections. Study of color intensified, sincehave been identified in fungi. Consequently, the late 19th century, to understand:diversity of fungi pigments is the second in importance,after plant flavonoids. Fungi are not photosynthetic C The phenomenon for survival of animals and plants,and do not contain chlorophyll. Carotenoid C The relation between color and evolution theories;distribution in fungi is restricted to some orders (e.g., andPharagmobasidiomycetidae, Discomycetes). In addition, C The role imparting in comparative physiology.flavonoids are scarce in fungi whereas riboflavin impartsthe yellow color in the genera Use of natural colorant in Thus, studies on biocolorant are greatly impulsed byfood industry appears to have multidimensional potential their multiple functions [27]. The art of coloring spread[22]. For example, in addition to coloring property,- widely with the advancement of civilization [28].carotene may be used in food as an essential vitaminsource or betalains as source of essential amino acid or Source of Biocolorants: Plants, animals andanthocyanins as quality control marker of food stuffs. microorganisms are the sources of natural biocolorants,Flavonoids are colorants with high pharmacological but few of them are available in sufficient quantities forpromise. Sometimes, diets with carotenoid mixtures are commercial use as food colorant and mostly are plantrecommended instead of having just one particular origin. For biotechnological production of such colorants,carotenoid, because great variability of radicals and plants and microorganisms are more suitable due to theirmicroenvironments take place in vivo [23]. Process understanding of proper cultural techniques anddevelopment and use of new and available technologies processing. Natural colorants obtained from plant originfor production optimization are the important area of are pepper, red beet, grapes, saffron [29, 30]. Nowadays,research on natural food grade colors. Therefore, it fermentative production of food grade pigments arerequires a thorough review of the biotechnological available in the market, for example; color from Monascuspotentials of ‘natural food grade biocolorants’. sp., astaxanthin from Xanthophyllomyces dendrorhous,

History of Biocolorants: Man has always been interested from Ashbya gossypii and -carotene from Blakesleain colors. The art of dyeing is of long past and many of trispora. Also a number of microorganisms producethe dyes go back into prehistory. In Europe, it was biocolors in good amount that includes Serratia andpracticed during the Bronze Age. The earliest written Streptomyces [31].record of the use of natural dyes was found in Chinadated 2600 BC. In Indian subcontinent, dyeing was known Biocolorants in Food Industry: To consider the naturalas early as in the Indus Valleyperiod (2500 BC) and has biocolors as the colorant, stability, yield and price arebeen substantiated by findings of colored garments of mostly the constrains. Most of them are sensitive to pH,cloth and traces of madder dye in the ruins of heat and sunlight [32]. Inspite of such factors, the naturalMohenjodaro and Harappa civilization (3500 BC). In biocolorants are gaining importance because of healthEgypt, mummies have been found wrapped in colored and hygiene, nutrition, pharmaceutical activities, fashioncloth. Chemical tests of red fabrics found in the tomb of and environmental consciousness, indicates relativeKing Tutankhamen in Egypt showed the presence of dependency on natural products. Colors derived fromalizarin, a pigment extracted from madder. minerals (lead chromate, copper sulphate) may cause

The cochineal dye was used by the people of Aztec serious health problems [33]. However, in last fewand Maya culture period of Central and North America. decades, synthetic additives are severely criticized andBy the4th century AD, dyes such as woad, madder, weld, consumers show inhibition toward these products,Brazilwood, indigo and a dark reddish-purple were consequently they prefer to use the natural colorantsknown. Brazil was named after the woad found there [24]. [35]. In the 1960s in US, the environmental activistsHenna was used even before 2500 BC, [25] while saffron demonstrated against the use of such food additives

Arpink red color from Penicillium oxalicum, riboflavin

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and this attitude was spread out widely. Activists also in cosmeticsand foods, particularly in dairy productscampaigned for the natural colorants highlighting their (cheese), confectionary (soft and hard candy), fish andnutritional characteristics as a sales tool. A strategy that meat products, fruit products, beverages, snacks, beerhas failed earlier, later converted it into a total success, and wine. It is more stable in photo degradation than thatresulting from changes in social attitude. Thus, a of carotene [42]. However, canthaxanthin is notworldwide tendency to use natural colorants is generated. considered as food additive under EU regulation.Currently, people interpret the content of synthetic Cochineal insect (Dactylopius coccus), best cultivatedproducts as contaminant and the tendency has been on Opuntia ficusindica, extract is known to have carminereinforced [36]. There are number of advantages of using or carminic acid, which appears as magenta-red uponnatural biocolorants, over synthetic colorants, further application. Water insoluble forms of carmine exhibit aboosted because of their pharmacological properties. color range from pink to purple. It is resistant to light,Products are of good market value if they are colored with heat and chemical oxidation, often it is more stable thannatural compounds, for example; annatto in Cheddar synthetic food colorants but unstable at low pH. Thecheese [37]. water soluble form is used in alcoholic drinks with

Food Grade Natural Biocolorants: Natural biocolorants variety of products. Together with ammonium, carmine isare under ‘exempt from certification’category of FDA and used inmeat, sausages, processed poultry products,EU for food use and are: annatto extract, dehydrated red alcoholic drinks, beverages, bakery and dairy products,beet, canthaxanthin, carotene, Dactylopuis coccus extract, including desserts and sweets. On an average, peoplecotton seed meal, grape skin extract, fruit and vegetable consume one or two drops of carminic acid annually withjuices, tagetes extract, carrot oil, oil of corn endosperm, food [43]. Cotton seed meal is the by-product after oilpaprika and paprika oleoresin, riboflavin, saffron, turmeric extraction from cotton seed. Gossypol is a fat solubleand turmeric oleoresin and xanthophylls (flavoxanthins, yellow pigment that occurs in cotton seed in bound andrubiaxanthins, zeaxanthin). Chlorophylls, in spite of its free forms. The bound form of gossypol is combined withlarge intake amount per day. Yellow to orange color of free amino acids which constitute a medium qualityAnnatto comes from the outer layer of seeds of the protein feed [44]. Grape skin extract (enocianina) impartstropical tree Bixa orellana. The carotenoids, bixin and a reddish purple color to beverages [45]. Some fruitsnorbixin, are responsible for appearance of yellow to contain a single type of anthocyanin (cyanidin in apple,orange color. The pH and solubility affect the color hue; cherry, fig, etc), some contain two major types (cyanidingthe greater the solubility in oil, the brighter is the color. and peonidin in ascherry, canberry); or some with severalWater soluble, oil soluble and oil/water dispersible forms anthocyanins (grape). As colorant, grape juice is availableof annatto are available. Since it precipitates at low pH, is in a variety of colors: red (shades of cherry, raspberry oralso available as emulsion, an acid proof state [38]. strawberry), purple and yellow. It gives a strawberryredAnnatto has been used for over two centuries as a food shade in application. It is used to color a numbercolor especially in cheese and in various other food ofnon-beverage foods, including gelatin desserts, fruitproducts [39]. Red beet (Beta vulgaris) extract shows fillings and in certain confectionaries. Anthocyanins fromvariety of colors, depending on their content of yellow banana bract and Oxalis triangularis are found ascompound and may have a good flavor. Also a potential source of food colorants [56]. Acylatedbluish-red color produced by a compound known as anthocyanins from different edible sources (black carrotbetanin and is stable at higher pH range than red cabbage cultivars) are also used in food industry [47]. Vegetableextract [40]. There is no limit on its upper usage level. It juice is actually fermentable liquid product, eitherhas wide application in different food commodity from unfermented or lactic acid fermented, obtained from thebeverages to candy and from dairy to cattle products [41]. edible part of one or more vegetables for directCanthaxanthin, a carotenoid, is commercially produced consumption and preserved exclusively by physicalfrom the algae Haematococcus lacustris. Canthaxanthin means. The juice shall be free from skins, seeds and otheris known mainly as the natural pigment of the coarse parts of the source vegetables. Tomato juice andorangeyellow chanterelle mushroom. Canthaxanthin has blends based on tomato have long been popular andvarious physiological functions and can be converted account for over 90% of the non-fruit juice trade.into vitamin A under stress. Canthaxanthin is used in Lycopene is the principal compound derived from tomato,poultry for the appearance of color shade of the yolk, enlisted under US system. Under EU legislation,

calcium carmine; but water insoluble form is used in a

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lycopene (E 160d) is considered as food additives. It is Carotenoids: Carotenoids are one of the most importanthighly stable under a wide range of temperature and groups of natural pigments. It has been estimated thatpH, hence used as common food colorant. It is available nature produces about 100 million tons of thesepigmentsin liquid form or as cold-water dispersible powder. Carrot per year. They give brilliant yellow and red colours tohas long been a component of tomato blends [13]. many fruits, vegetables, roots, flowers and autumn leaves;Red-cabbage juice produces a bright pink to red color they produce the colour of the egg yolk, of many algae,to a product with a pH less than 4.0 and is soluble in yeasts and mushrooms, Crustaceae as well as of thewater, but not in oil. Use of Marigold (Tagetes erecta) feathers and skins of many birds. The carotenoids haveflowers as source of food colorants is known from attracted the curiosity of scientists since the beginningAztec civilization. Marigold flowers are by far the most of organic chemistry, actually since 1818. Numerousabundant natural source for commercial lutein [48]. The outstanding chemists made valuable contributions andantimutagenic activity of carotenoids of Aztec marigold many tools and methods have been introduced intowas evaluated. The presence of some essential amino organic chemistry via the carotenoids'. Imention the earlyand fatty acid increases the nutritional quality of carrot analyses (by Willstätter and Zechmeister), theseed oil. Corn endosperm oil is a reddish-brown liquid rediscovery of column chromatography by Kuhn andcomposed chiefly of glycerides, fatty acids, sitosterols Karrer and of the thin-layer chromatography by Callebautand carotenoid pigments obtained by isopropyl alcohol [l2], the elucidation of the isoprene rule by Ruzicka andand hexane extraction from the gluten fraction of yellow of the symmetrical structures of squalene, /3-carotenecorn grain [49]. It is used in chicken feed as color and lycopene by Karrer, the studies on comparativeadditive.Paprika (Capsicum annuum) is the pioneer and biochemistry by Goodwin3, the first rules onwidely [50] used carotenoid as food colorant. In paprika, u.v.absorption of polyenes by Richard Kuhn, thered carotenoidsare dominated by canthaxanthin, investigation of cis-trans isomerism [54] and the firstcapsorubin where as yellow xanthophylls includes syntheses by Karrer and Inhoffen. The application ofcryptoxanthin, zeaxanthin, antheraxanthin and -carotene n.m.r. spectroscopy by Jackman and Weedon5 proved to[51]. Paprika oleoresin is mainly extracted from the pods. be a valuable tool for the identification of carotenoids,It contains three main naturally occurring pigments: the synthesis of which was especially advanced by thecapsanthin, capsorubin and carotene. This combination development of condensation reactions for the buildupproduces a bright orange to red-orange in food products. of the conjugated chain (Wittig6, Homer7). We aimed atThe oleoresin is oil soluble, when emulsified becomes the development of commercial syntheses of somewater dispersible [52]. Riboflavin (vitamin B2) has a selected carotenoids arid the investigation of their use asvariety of applications as a yellow food colorant. Its use food colourants. By substituting natural colouring mattersis permitted in most countries. Applications include for artificial dyes we fulfil a physiological desideratum thatdressings, sherbet, beverages, instant desserts, ice good food should not contain unnatural pigments [54].creams, tablets and other products. Riboflavin has aspecial affinity for cerealbased products, but its use in Production of Carotenoids from Natural Sources: Thethese applications is somewhat limited due to its slight classical method for producing /3-carotene and otherodour and naturally bitter taste. carotenoids is solvent extraction of the plant material.

Other Application in Food Industry provitamin A activity of the extract and the degree ofFood Preservatives: Several natural biocolorants, pigmentation of eggs and broilers, after feeding, areincluding anthocyanin, additionally show antagonistic indicated.activity to certain bacteria, viruses and fungi thus to Carrots, palm oil, alfalfa, dried grass and leaves areprotect food from microbial spoilage (Bridle and [53]. the starting materials for practically all preparations ofSome are also active against protozoa (Leishmania natural carotene with provitamin A activity. The extractsbrasiliensis) and insects (Calliphora erythrrocephala). from alfalfa, grass, leaves, Tagetes and yellow maize,Sometimes, carotenoids can act as sun screen to containing either lutein or zeaxanthin, are valuablemaintain the qualityof food by protecting from intense supplements to chicken feed because allxanthophyllslight. It has beenreported that corn carotenoids inhibit colour the egg yolk in contrast to the carotenes. Besidesthe synthesis of aflatoxin by Aspergillus flavus (90%) /3-carotene, annatto which is an extract of the seeds ofand by most of the A. parasiticus (30%) strains [54]. Bixa Orellana is also used for colouring dairy products.

The natural source, the main pigments, possible

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For red carotenoids large quantities of paprika are (e.g., turmeric) could be made in this way. Colorantsextracted. Torula yeast and certain algae are other sources made in this way will probably face some hurdles, at leastof red polyene pigments used as colourants for oil and in Europe where genetically modified food is generallycheese. The market for natural carotenes has declined viewed with a large degree of skepticism by thesince the introduction of synthetic carotene. Today, only consumers. Finally, new colorants would have to beone crystalline carotene preparation extracted from approved by the authorities, which is very costly becausedehydrated carrots is still on the market. Recently it was of the various toxicological studies needed to confirmannounced that /3-carotene and further carotenoids will the safety of a new food additive. The above gives a fairlybe produced in Brazil by fermentation from mated strains compressed overview of the most important naturalof Blakeslea trispora [54]. food colorants. The reader is referred to a number of

CONCLUSION pigments and food colorants [12-16].

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