Trends in Biosciences 10(21), Print : ISSN 0974-8431, 4057-4064, 2017

REVIEW PAPER Natural Pigments as Potential Food Colourants: A Review

MANJUNATH J. SHETTY, P. R. GEETHALEKSHMI AND MINI. C Department of Processing Technology, College of Agriculture, Vellayani, Kerala Agricultural University *email: [email protected]

ABSTRACT History Palatability of food is enhanced by attractive colours. The earliest written record of the use of natural dyes Colour, more than any other factor, influences the dates back to 2600 BC in China and addition of colourants acceptance of products by consumers. Colour additives to foods is reported in Europe during the Bronze Age. It is have been used since time immemorial to enhance the also reported that around 1500 BC in Egyptian cities candy aesthetic value of food. Synthetic food colourants are being makers used to add natural extracts and wine to improve recognized to be carcinogenic and harmful to the the appearance. The first synthetic colour (mauvine) was consumers, hence plant pigments are the alternative. developed by Sir William Henry Perkin way back in 1856. Pigments produce colours that we observe at each step of The beginning of the 19th century was remarked for the our lives and plants are the principal producers. Several bulk of production and recovery of synthetic colours from plants are known to yield red pigments, blue pigments, the petroleum derived products like aniline, therefore they brown pigments and yellow-orange pigments which can be were called ‘coal-tar’ colours because the starting materials used for food colouring. Regulating the use of synthetic were obtained from coal (Burrows, 2009). colour and use of natural food colours are promising to Food colourants have additional health benefits. Substances which add or restore colour in Key words Pigments, Synthetic, Colourants, a food, that includes natural/synthetic constituents which are normally not consumed as foods as such (FSSAI, Carcinogenic, Natural 2011). Food colours can be classified into three groups; Colour of a food substance is important to indicate natural food colourants, which refer to ones that are its freshness and safety that are also indices of good synthesized naturally; nature-identical colorants, which aesthetic and sensorial values. In the recent years, although synthesized in industries, mime the natural ones colouring of food with pigments produced from natural and finally the artificial/synthetic colourants (Msagati, sources is of worldwide interest and is gaining importance. 2013). Natural & synthetic food colour market is projected These pigments are looked upon for their safe use as a to reach $2.3 billion by 2019 growing at a rate of 4.6% natural food dye in replacement of synthetic ones because annually. of undesirable market. A well textured food, rich in nutrients Reasons to use food colourants and flavour, cannot be eaten unless it has the right colour. Nowadays, people know that foods have their own Colour additives have long been used as a means of natural colours, but most people live in big cities, far from enhancing the aesthetic value of foods, beverages and the places of food production, and processing and cosmetics and for identifying drugs and other products. transportation of foods are required. During such events, The demand for natural source of such compounds is foods may lose some of their properties and some additives increasing day by day because of the awareness of positive must be added to permit such products having desirable health benefits out of natural compounds. It is therefore, appearance and are safe for consumption. Moreover, it is essential to explore various natural sources of food grade possible to enumerate the reasons to use food colourants colourants and their potentials. An attractive colour is often (Shamina et al., 2007). associated with quality and freshness. Consumers have 1. To restore the original food appearance expectations with regards to the typical colour of certain foods. A non-attractive colour makes a product look as no 2. To intensify colours that are normally found in food longer fresh and likely to be rejected. Colour has important and the consumer will associate this improved colour psychological implications, particularly true in case of food. with food quality. The initial perception of food by consumer is visual. 3. To protect the flavour and light susceptible vitamins. Therefore colour has a very important role and the 4. To preserve the identity or character by which food acceptance or rejection of a food often depends on it. is recognized. 4058 Trends in Biosciences 10 (21), 2017

Classification of food colourants and . The reddish-purple colour of most common fruit and berries is produced mainly by a) Synthetic colours: These do not occur in nature and pigments. On a global basis, a few carotenoids, are produced by chemical synthesis. Seven synthetic two , the predominant green and colourants, belonging to four distinct chemical about three seasonally occurring are the most classes, are permitted by the FDA for use in foods abundant pigments in plants and algae (Hendry, 1996). and in orally ingested drugs and cosmetics: FD&C , some less-common carotenoids and flavonoids Red No. 40, FD&C Red No. 3, FD&C Yellow No. 5, as well as all of the other minor pigment classes. It is FD&C Yellow No. 6, FD&C Blue No. 1, FD&C Blue estimated that 4000 tonnes of anthocyanin are consumed No. 2 and FD&C Green No. 3. Citrus Red No. 2 is each year in the UK, mainly from fruits (Henry, 1996). Several permitted only for colouring skins of oranges and plants are known to yield red pigments (Bixa orellana, Orange B for surfaces of casings of frankfurters and Beta vulgaris), blue pigments (Vitis vinifera, Vaccinum sausages. They share the common property of water mytillus), brown pigments (Camellia sinensis, Lawsonia solubility, being conferred by the presence of one or inermis) and yellow-orange pigments (Curcuma longa, more sulphonic or carboxylic acid groups (Parkinson Tagetes erecta) which can be used for food colouring and Brown, 1981). (Lakshmi, 2014). b) Nature-identical Colours: These colours are also Animals and microorganisms are also the sources of manufactured by chemical synthesis, but do not natural pigments, but few of them are available in sufficient require FDA certification and are considered quantities for commercial use as food colourant and mostly chemically and functionally indistinguishable from are plant origin (Haliwell, 1996). the same colourant found in nature. c) Natural Colours: Natural food colour is any dye, Major classes of natural pigments pigment or any other substance extracted using conventional methods, from vegetable, animal, mineral Anthocyanins are natural colourants belonging to or other sources capable of colouring food drug, the family of secondary metabolites; they are a cosmetics or any part of the human body; colours diverse group of intensely coloured pigments synthesized come from a variety of sources such as seeds, fruits, in plants responsible for the often brilliant purple and blue vegetables, algae and insect. colours in fruits, vegetables, flowers, leaves, roots and other Pigments plant storage organs. They are found predominantly in outer Pigments produce the colours that we observe at each cell layers such as the epidermis and peripheral mesophyll step of our lives, because pigments are present in each one cells. They are water soluble, facilitating their easy use in of the organisms in the world, and plants are the principal aqueous media as food colourants (Markakis, 1992). producers. They are in leaves, fruits, vegetables, and Anthocyanins are allowed as a food additive flowers; also, they are present in skin, eyes, and other animal (designated E 163). Anthocyanins from tart cherries (Prunus structures; and in bacteria and fungi. Natural and synthetic cerasus L.) have been stabilized with phosphoric acid, pigments are used in medicines, foods, clothes, furniture, maltodextrin and a- and b-cyclodextrins (Chandra et al., cosmetics, and in other products. (Frick and Meggos, 1988; 1993). The colour is susceptible to temperature, oxygen, Hari et al., 1994). Foods particularly fruits and vegetables, UV light and acidity. At different pH, anthocyanin reveals are naturally coloured mainly by four groups of pigments: several colours. For pH indication, it acts as natural source. the green chlorophylls, the yellow-orange-red carotenoids, 1 ml of the anthocyanin extract (methanol: HCl) of seedless the red-blue-purple anthocyanins and the red grape peel was added in test tubes. From the pH of 1to 14, (Mittal et al., 2007). Because of the deficiencies of existing the extracts was adjusted using the acid as 1N HCl and the natural food colourants, the demand for natural pigments base as 1N NaOH by the help of pH meter. Finally, the is repeatedly raised by the food industry. This demand can extracts were mixed well to detect the colour. The colour be fuelled by research to offer a more natural-healthy way changes were recorded (Bondre et al., 2012). Colour loss of colouring foods. may be prevented by storing at low temperature in dark Natural distribution of pigments containers or under oxygen free packaging. The highest-seed coat of black soybean-2,000mg / Among the biological sources, many natural dyes/ 100g. Other sources include apple, berries (blackcurrant, colourants are obtained from plants, producing various boysenberry, blueberry, strawberry, blackberry, raspberry, colours such as red, yellow, blue, black, brown and cranberry, elderberry, loganberry, chokeberry etc.), black combinations of these. The wide range of green colours in carrot, cabbage, cherry, grape, radish, red onions and sweet leafy vegetables is produced by different combinations of SHETTY et al., Natural Pigments as Potential Food Colourants: A Review 4059

Table 1. Major classes of pigments Latin American cuisine to add a deep red colour as well as distinctive flavour notes to fish, meat and rice dishes. In Classes of natural pigments Sources the United States, annatto extracts are primarily used to impart orange/yellow hues to cheese and other dairy foods. 1. Anthocyanins (Red to Blue) Blue grape, blueberry, The annatto tree, B. orellana, is a large, fast-growing shrub cherry plum, kokum fruit, Hibiscus, cherry, of the tropics. The large clusters of brown or crimson apple and Jamun capsular fruit-containing seeds coated with a thin, highly coloured resinous coating serves as the raw material for 2.Carotenoids (Orange yellow Annatto seeds, alfalfa, to Red) carrot, palm oil, saffron the preparation of the colorant annatto, which is prepared and marigold by leaching the annatto seeds with an extract prepared from one or more food grade materials including various solvents, 3.Betalains (Red, Purple) Beetroot, cactus, amaranths edible vegetable oils and fats and alkaline aqueous and alcoholic solutions. The chief colouring principle found in 4. Safflower florets the oil-soluble extracts of annatto is the bixin; 5.Chlorophylls (Green) Green plants the major colorant in the alkaline aqueous extracts is norbixin (Marmion, 1991). 6. Lycopene Tomato, watermelon Clinical application: Carotenoids are photo-protectants. 7.Miscellaneous: Lee et al. (2000) suggested that supplementation with ii. Carminic acid (Red) Cochineal insects natural carotenoids may partially protect human skin from UV-A and UV-B induced erythrema. Astaxanthins, a iii. Caramel Modified sugars carotenoid widely used as an intermediary in reproductive (Fergus and Reuter, 2004) process. Betalains potato (Raghavendra et al., 2011). Khan and Giridhar estimated annual production Clinical application: Enhancement of sight acuteness potential of plant betalains from edible sources (red (Timberlake and Henry, 1988), treatment of various blood beetroot, Swiss chard petiole, cactus pear fruit, pitaya fruit circulation disorders resulting from capillary fragility (Wang and amaranth seed); production of beetroot far exceeded et al., 1997), radiation protective action (Akhmadieva et al., those of the other sources. Betalains are a group of red and 1993), to examine the anti-cancer effects (Stoner et al., 2007). yellow colour water soluble pigments found in red beet, Carotenoids cactus fruits and in some flowers. They are stable to pH range 4-6 but degraded by thermal processing. The Carotenoids are group of responsible sensitivity of betalains to different factors suggests that for yellow, orange and red colours of foods and having an their application as food colourants is limited. Based on important role in the prevention of human diseases and these properties, betalains can be used in foods with a maintaining good health. In addition to being potent short shelf-life, produced by a minimum heat treatment, and antioxidants some carotenoids also contribute to dietary packaged and marketed in a dry state under reduced levels vitamin A. Although the chemistry of carotenoids has been of light, oxygen, and humidity (Rayner, 1993; Von Elbe, 1977) studied extensively, their bioavailability, metabolism and biological functions are only now beginning to be Clinical application: antiviral and antimicrobial investigated (Rao and Rao, 2007). activities, In some places in Mexico, an infusion of Bougainvillea bracts mixed with honey is used widely for ß-, ß-apo-8-carotenal and canthaxanthin: cough. Reduces cardiovascular disease and diabetes, These are the most commercially important of carotenoids. Reduces risk of cancerand signs of aging (Anti-aging) and Beta-Carotene is the isomer of the naturally occurring prevents of chronic diseases. carotenoid, carotene. Carotene is the pigment largely responsible for the colour of butter, cheese, carrots, alfalfa Chlorophyll and cereal grains. The colourant is synthetically extracted Chlorophylls have been the least studied of the food with acetone, which results in the all-trans form. Both the pigments. Chlorophylls (E 140) are vegetable pigments that synthetic and natural sources of ß-carotene are permitted occur naturally in plants and confer colour. Among the five colour additives (Marmion, 1991). different chlorophylls that exist, only two (a and b) are used Annatto in the food industry as colourants. Their complex structure is difficult to stabilize, being this the main drawback of their Annatto is a natural food colourant extracted from use in the industry, which has studied mechanisms of the seeds of the Bixa orellana L. plant. Annatto is used in 4060 Trends in Biosciences 10 (21), 2017 retaining or replacing the magnesium ion within the 25 mg; 50% of this is in the form of processed tomato structure. The used commercial colourants of chlorophylls products. are extracted from alfalfa, and have been employed in dairy Clinical applications: Lycopene has attracted attention products, soups, drinks and sugar confections because of its biological and physicochemical properties, (MacDougall, 2002). especially related to its effects as a natural antioxidant (Sies Clinical applications: Chlorophyll also helps support et al., 1998). The consumption of tomato products has the healthy flora in the bowels which also help improve our shown promise for the prevention of prostate cancer. immunity function. Chlorophyll has great antioxidant Miscellaneous pigments capacity and therefore it helps neutralise free radicals and limit oxidative damage within the body. From several animal Carmine and in vitro research studies, chlorophyll and its derivatives Carmine, also called Crimson Lake, Cochineal, Natural have been shown to possess anti-carcinogenic properties Red 4 is a pigment of a bright red colour obtained by aqueous which mean they have the ability to help prevent or slow extraction of cochineal, which is derived from the dried down cancer. Studies have also found that chlorophyll also bodies of the gravid female insect Coccus cacti, and is improves the health of blood cells by increasing the uptake used as a general term for a particularly deep red colour of oxygen in the blood. Austrian scientists have found that (Lucas et al.,2001). Although principally a red dye, it is chlorophyll and its derivatives successfully slow down the found in many foods that are shades of red, pink and purple. oxidation of LDL (the ‘bad’ cholesterol) which is a major Since typically it is used in low quantities, the likelihood of contributor to cardiovascular disease. sensitization to carmine ingestion is low and is more likely Anthoxanthins to occur through high levels of exposure through occupational or cosmetic exposure. But it has been known Anthoxanthin is a pigment and is similar to to cause severe allergic reactions and anaphylactic shock anthocyanin, but it exists in less oxidized state as the in some people (Green field, 2005). Food products oxygen on the central group is uncharged. It is actually a containing a carmine-based food dye may prove to be a composite of compounds known as flavones, and concern for people who are allergic to carmine. . Anthoxanthins are white, pale yellowish, water soluble pigments found in a plant’s cell sap. They contribute Caramel the cream and white colour of cauliflower, onions, white Caramel colour or caramel colouring is a water soluble potatoes and turnips. Short cooking is desired. With food colouring. It is made by a careful controlled treatment prolonged heat, the pigment turns into a brownish grey of carbohydrates, in general in the presence of acids, alkalis colour (Clifford and Cuppett, 2000). or salts, in a process called caramelization. It has odour of Clinical applications: Anthoxanthins are thought to burnt sugars and somewhat bitter. Its colour ranges from have health-promoting properties due to their high pale yellow to dark brown. Caramel colour is one of the antioxidant capacity in both in vivo and in vitro systems. oldest and most widely used food colourings, and is found The functionality in human health is supported by the useful in Beer, Brown bread, Chocolate, Cookies, Spirits ability of the pigments to induce human protective enzyme and liquor, Dessert mixes, Doughnuts, Fruit preserves, Ice systems, and by a number of epidemiological studies creams, Soft drinks (colas), Sweets (EFSA, 2011). suggesting protective effects against cardiovascular Application of colours in food diseases, cancers, and other age-related diseases (Cook Effects of natural colours on food products and Samman, 1996). Application of Amaranthus betacyanins in jelly, ice Lycopene cream, and beverage was evaluated in comparison to Lycopene is the pigment principally responsible for commercial colourants (Cai and Croke, 1999). The the characteristic deep red colour of ripe tomato fruits and betacyanins in these model foods exhibited brighter colour tomato products; it is also found in watermelon, papaya, characteristics than red radish anthocyanins, with similar pink grapefruit and pink guava. Processed tomato products colour stability during 20-week storage. The viability of are more available dietary sources of lycopene than fresh acylated anthocyanins from red radish, red cabbage, black tomatoes. Lycopene is a member of the carotenoid family; it carrot and grape skin extract to colour dairy products such is a natural fat-soluble pigment found in certain plants and as yogurt and sour cream having pH levels around 4.2-4.5. microorganisms, where it serves as an accessory light- (Giusti and Wrolstad, 2000). gathering pigment and to protect these organisms against Roselle anthocyanins were evaluated as natural the toxic effects of oxygen and light (Rao and Agarwal, colourant for hard candy. To determine the more suitable 1999). The average daily intake of lycopene is approximately concentrations of these natural colourants, the SHETTY et al., Natural Pigments as Potential Food Colourants: A Review 4061

Table 2. Natural pigments and their uses in food stuffs

Pigments Foodstuffs References

Annatto Cakes, Biscuits, Dairy products, Flour, Soft drinks, Snacks Hendry and Houghton, 1996,Rao et al., 2005, Scotter, 2009

Carotenoids Meat products, Sauces, Marmalades, Spice blends, Beverages, Baines and Seal, 2012 Milk products

Anthocyanins Confectionary, Fruit preparations Hendry and Houghton, 1996, Baines and Seal, 2012

Chlorophylls Dairy products, Soups, Drinks MacDougall, 2002

Curcumin Yoghurt, Baked goods, Dairy products, Ice creams, Salad Hendry and Houghton, 1996, dressings MacDougall, 2002

Carmine Jams, Gelatins, Baked goods, Dairy products, Non-carbonated MacDougall, 2002 drinks encapsulated pigment was used at levels ranged between temperature without any effects on the other organoleptic 0.05 to 0.3 %. Control sample was coloured with 0.019% attributes. (Hadyet al., 2014) synthetic colourant (Raspberry red, E124). Candy samples The incorporation of astaxanthin oleoresin to milk were sensory evaluated for their quality attributes i. e. colour; with different fat content, simulating the red-orange colour flavour, mouth feel, and overall acceptability. The results that can impart apricot colour (Mezquita et al., 2015). showed that colour of control sample, which coloured with Commercially important natural food grade dyes were synthetic dye scored the highest values for colour while evaluated in terms of their sensitivity against gamma those coloured with low levels of roselle anthocyanins i.e. ionizing radiation. Cochineal, annatto and turmeric dyes 0.05 and 0.1% were significantly different. However, adding with suitable concentrations were subjected to increasing roselle anthocyanis at a level of 0.2% was effective to doses up to 32 kGy and was analyzed by spectrophotometry produce hard candy with colour comparable to that of and capillary electrophoresis. The results showed different control sample. No significant differences could be found pattern of absorbance versus absorbed dose for the three between control sample and samples coloured with different systems (Helio et al., 2016). levels of anthocyanins regarding the flavor and mouth feel Regulatory aspects properties. (Selim, 2000) Regulatory aspects of food colouring in India (FSSAI, The studies on impact processing conditions on the 2011) stability of annatto dye incorporated in some baked and fried snack foods indicates that high loss of colour in fried Unauthorized addition of colouring matter prohibited. items as most of the added annatto leached out into oil Extraneous addition of colouring matter to be (Prabhakara et al., 2005). The anthocyanin-rich waste (ARW) mentioned on the label. in purple corncobs has been characterized to find a suitable Natural colouring matters which may be used are application in a food matrix. ARW provided milk with an – Beta-carotene attractive purple hue. ARW shows potential as a natural colourant for a pH range unusual for anthocyanin – Chlorophyll applications (Zing and Giusti, 2005). – Riboflavin The eggs of female quail fed with diets supplemented – Caramel with natural carotenoid (alfalfa (or Lucerne) concentrate, – Annatto tomato powder and marigold extract) showed an increase – Saffron in the yolk concentrations of lutein, zeaxanthin, lycopene – or turmeric and ß-carotene (Karadas et al., 2006). Purple carrot puree have a high concentration of acylated anthocyanins made Addition of inorganic colouring matters and pigments it more stable and suitable for mixing with strawberry during prohibited. jam processing. Adding up to 20% of purple carrot puree Use of permitted synthetic food colours in or upon during strawberry jam processing enhanced the colour any food other than those enumerated is prohibited. stabilization during and after six month of storage at room Maximum limit of permitted synthetic food colours 4062 Trends in Biosciences 10 (21), 2017

should not > 100ppm. Future prospects Colours used should be pure and free from any Consumer demands and their preferences for harmful impurities. naturally-derived colourants have increased exponentially, Authorities which are regulating food colours in widely associated with the image of healthy, safe and good different countries quality products, which constitutes a great challenge to food industries and related food science research Canada : Health Canada institutions (Carocho et al., 2014; Dias et al., 2015; Shahid Mexico : Norma Official Mexicana (Official Mexican et al., 2013). Global trend towards the use of natural Standard), abbreviated as NOM products in daily life opens new avenues for the increasing Australia : Food Standards Australia and New use of natural dyes and colours. Despite centuries of Zealand interest in natural pigments, our knowledge of their China : National Food Safety Standard distribution, availability and properties is limited. Japan : Japan External Trade Organization (JETRO) Worldwide, 70% of all plants have not been investigated. Therefore, novel plant pigments must be searched for Korea : Food regulation is under the Ministry of unprosecuted land or in the sea. Detailed study and Food and Drug Safety (MFDS) scientific investigation are needed to assess the real Middle East and Northern Africa: EU regulations in potential and ability of natural dye yielding general plants.Standardization of extraction methods for higher Advantages of natural food colours colour yielding plants. Need based use of colours in different food preparation should be evaluated. Improves the health condition of the consumer Non allergic and eco friendly LITERATURE CITED Better bio degradability and higher compatibility with Akhmadieva, A. K., Zaichkina, S. I., Ruzieva, R. K., and the environment Ganassi, E. E. 1993. The protective action of a natural preparation of anthocyanin (pelargonidin-3, 5- They have high medicinal value diglucoside). Radio biologiia. 33(3): 433–435. It saves the energy Baines, D. and Seal, R. 2012. Natrural food additives, Application process is less costly ingredients and flavourings. Woodhead Publishers, Disadvantages of natural food colours Cambridge, UK, 450p. Seasonal production Bridle, P. and Timberlake, C. F. 1997. Anthocyanins as Variation in quality and purity from source to source natural food colour selected aspects. Food Chem. 58: 103-109. Availability in limited shades Burrows J.D. 2009. Palette of Our Palates: A Brief History of Low concentration in source material Food Coloring and Its Regulation, Comprehensive Rev. Difficulties in the extraction from sources Food sci. Food Saf. 8(2): 394-402. Most of them are sensitive to pH, heat and sunlight Cai, Z. Y. and Corke, H. 2005. Characterization and (Hallagen et al., 1995) application of pigments from plants of the CONCLUSION Amaranthus. Trends in Food Sci. &Tech. 16: 370– Nature is rich in colour and the majority of plant 376. pigments can be widely exploited for the colouring of food. Carocho, M., Barreiro, M. F., Morales, P., & Ferreira, I. C. F. As the awareness among people towards natural dyes and R. 2014. Adding molecules to food, pros and cons: a their therapeutic uses are increasing because of their review on synthetic and natural food additives. nontoxic or less toxic properties, fewer side effects. New Comprehensive Rev. Food sci. Food Saf. 13(4) : 377- sources of plant pigments need to be available in sufficient 399. quantities for industrial extraction. To meet the need and Chandra, A., Nair, M. G., and Iezoni, A. F. 1993. Isolation demand of growing population, emerging technologies and stabilization of anthocyanins from tart cherries such as high hydrostatic pressure, pulsed electric fields, (Prunus cerasus L.). J Agric. Food Chem. 41: 1062– sonication and membrane technologies could be potential 1065. methods for the enhanced extraction of pigments and Clifford, A. H. and Cuppett, S. L. 2000. Review: bioactive compounds from plant or animal materials in future. Anthoxanthins—nature, occurrence and dietary burden. Synthetic food colourants are being recognized to be J. Sci. Food Agric. 80: 1063– 1072. carcinogenic and harmful to the consumers, hence plant Cook, N. C. and Samman, S. 1996. Review: Flavonoids— pigments are the alternative. SHETTY et al., Natural Pigments as Potential Food Colourants: A Review 4063

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Received on 27-04-2017 Accepted on 17-05-2017