NATURES PALLETTE : EMRGING TECHNOLOGY ABOUT COLOUR

In recent years consumers are more concious about their health. They are mostly prefer natural as well as nutrious food. In this presentation i have included the content about natural colours which is extracted from the natural resources. by reading this presentation they will get clear information about natural and artificial colours.

1. Karthika Rajendran 2023-DDM-03 PhD Dairy Chemistry DC -691 Doctoral S eminar I nature’s PALETTE: AN EMERGING FRONTIER FOR C O L O U R I N G DAIRY PRODUCTS

2. Overview of presentation Introduction History Classification Sources Extraction techniques Natural sources in dairy products FDA Conclusion Dairy products Pigment yielding source Pigment extraction Extracted pigment Colored dairy products Labelling to indicate the presence of natural colors

3. According to FDA , A color additive is any dye, pigment , or other substance that can impart colour to a food , drug, or cosmetic. WHAT IS colOUr ? FDA, 2023.

4. Why COLOuRS ? Health Colour Global data consumer survey, 2022. 01. CONSUMERS ATTRACTION : To make food more attractive, appealing, appetizing, and informative.

5. To enhance colour that is already present 03 To colour the uncoloured food 04 To replace and restore colour lost during processing 02 Global data consumer survey, 2022.

6. H istory 19th The beginning of the 19 th century was remarked for the bulk production of synthetic colours like aniline. 2600 BC The earliest record on the use of natural dyes were found in china 2500 BC Dyeing was known in the indus valley period 2500 BC Saffron is mentioned in the bible 400 BC Egyptians coloured candy, and wine 1856 Sir william henry perkin developed first synthetic colour Susan, 1982 Sarkar et al., 2023. Natural colours synthetic colours

7. Types Beyer et al., 2002.

8. Natural colours Vs Synthetic colours Obtained from natural sources Processed by physical means Less stable Less bright Not uniform No health harm Good consumer acceptability High microbial contamination Expensive Obtained by chemical reaction High stability to light Highly coloured Color uniformity Health problems Consumer acceptability? Less costly Low microbial contamination Chávez et al., 2019

9. The natural colours have been classified 1. Plants 2. Animals 3. Microbes 4. Minerals Tripti Singh et al., 2023

10. animals 1. Cochineal ( Dactylopius coccus ): Native - South America and Mexico. It is a parasite which lives on cacti of genus Opuntia feeding on moisture and nutrients. The dye stuff extracted from this insect and its eggs is Carmine, which is red in colour. Carmine is used as a food dye in juices, ice cream, yogurt , and candy, and as a dye in cosmetic products such as eye shadow and lipstick. But as a food dye it has been known to cause severe allergic reactions. Lakshmi et al., 2014.

11. Pepsico - Red grapefruit drink Danone - Flavored yogurt (strawberry)

12. Wrigley – Candies The company substituted Red no 40 for carmine Red velvet cake mix contained cochineal extract

13. 2. Female Cuttlefish (Sepia officinalis L): It has rich concentrates of orange - red pigment in the accessory nidamental glands. The dye is called Sepia or squid ink. The sepia pigment is used in capsule printing ink which has been patented in European Patent Application EP1361258 . It is used in Spanish cuisine bread. Black pasta is often made using cuttlefish ink. Sepia officinalis L Female Cuttlefish

14. 3.Cephalopod is another member of the molluscan Cephalopod ink is generally obtainable from fishmongers or gourmet food suppliers and in cooking. it is used as a food colouring and flavouring , in pasta and sauces.

15. 4. Monascus purpureus : Monascus purpureus are fungus. Red pigments. The use of this colour additive is not yet regulated in the European Union, United States and Brazil, Philippines, Taiwan among other regions. Oriental countries such as Japan make extensive use of these pigments since decades - as water soluble pigments in candies or red pigment for red rice wine . It is also used as cholesterol-lowering agent.

16. Microbial source Riboflavin, Beta-carotene, Canthaxanthin , Prodigiosin , Phycocyanin , Melanin, Violacein , Astaxanthin , and Lycopene are the major pigments have been reported to produced from microoraganisms . Rana et al., 2021. Melanin is a natural pigment which is known to be produced by a wide variety of microorganisms such as Aspergillus fumigates (fungus), Vibrio cholerae (bacteria), Shewanella colwelliana (bacteria) , Alteromonas nigrifaciens (bacteria) . This pigment is also present in animals and plants . Uses - eye glasses, cosmetic creams, pharmaceuticals, they are also added in food items. The pigment is also reported to be associated with anti- HIV activity Sen et al., 2019 .

17. B2 vitamin Water soluble pigment Yellow colour Food additive in dairy products , sauces, baby foods, fruit, and energy juices. Candida guilliermundii (fungi), Debaryomyces subglobosus (yeast), Eremothecium ashbyii (fungi), Ashbya gossypi (fungi), Clostridium acetobutylicum ( bacteria) Riboflavin Beta-carotene Red-orange pigment Water insoluble Very good source of vitamin A Dunaliella salina(algae), Blakeslea trispora(fungi), Mucor circinelloides(fungi), Phycomyces blakesleeanus (fungi), Rhodotorula glutinis (fungi), Rhodotorula gracilis (fungi), Rhodotorula rubra (fungi). Orange to dark pink colored Lipid soluble pigment. Halobacterium sp. And Bradyrhizobium sp.(bacteria) Nannochloropsis gaditana (algae) Canthaxanthins are effective antioxidants and inhibit the oxidation of lipids. Canthaxanthin Dufossé , L. (2018).

18. Prodigiosin Red coloured Water insoluble Serratia marcescens (bacteria), Vibrio psychoerythrus , Rugamonas rubra , Streptoverticillium rubrireticuli yogurt, milk and carbonated drinks Insecticidal, antifungal, antibacterial, anticancer, and anti-malarial Blue coloured Water soluble It is found in Spirulina sp. It is used in sweets, ice creams and also as a dietary supplement rich in proteins. Act as bio-control agent that have anti-bacterial, anti-fungal and anti- alzhelmeric activity Phycocyanin Sen et al., 2019. Purple coloured pigment Water insoluble Chromobacterium violaceum , Pseudoalteromonas , Collimonas , Janthinobacterium , Cosmetics, food, medicine and textiles Antibacterial, anticancer, antiviral, antiulcerogenic , Violacein

19. Approved meat colouring agent in several countries. Water insoluble biopigment belonging to carotene. It is present in tomato and other red fruits and vegetables. E. Coli Yeasts fungus Lycopene Red coloured extracellular metabolite of the anthraquinone class produced by Penicillium oxalicum (fungus). It is suggested to have anticancer effects when used as food supplements Arprink red They are red ( monascorubramine and rubropunctamine ), yellow ( ankaflavin and monascin ) and orange ( rubropunctatin and monascorubrine ) colored pigments M. pilosus , M. purpureus , M.ruber and, etc. are being used as food colourants for many years in red wines, yogurt, sausages, and Meats. They are also known to exhibit antimicrobial, anticancer, anti-obesity, and antioxidant activities MONASCUS Rana et al., 2021.

20. NATURAL COLORS -PLANTS 1. Cornflower ( Centaurea cyanus ): Antioxidant, antibacterial and astringent 2. Indigofera tinctoria : Antihyperglycemic activity, antioxidant, Anti-inflammatory activity, antibacterial, antidiabetic activity and anticonvulsive agent 3. Bilberry ( Vaccinium myrtillus ): The fruit juice is red in colour and this turns blue in basic medium Lower blood sugar in diabetic patients. 4. Blueberry( Sambucus nigra ): Flowers are used for medicine, Fruits as dyes for basketry, arrow shafts, flute, whistles, clapper sticks. Elderberries are high in vitamin C. Purple to blue color : Centaurea cyanus Indigofera tinctoria Vaccinium myrtillus Sambucus nigra Rana et al., 2021.

21. Red color: 1. Annatto: Gloucester cheese-16th century Cheshire, red leicester cheese and cheddar - scotland . In spanish it is called as local saffron Yellow to orange color Bixin (fat soluble) and norbixin (water soluble) 2. Betalain Yogurts, ice cream Antioxidant, antimicrobial and antiviral activity 3. Paprica : Mixture of capsanthin and capsorubin , Both are carotenoids, Red color Cosmetics and medicine Beeta vulgaris Paprica

22. Yellow - orange COLOR Turmeric : It is called “ the golden spice of life. It has been considered has the poor man’s saffron. It is used as an alternative to saffron. Curcumin is the primary pigment of color. Dairy products , beverages, cereal, pickels , sausages, Confectionaries, ice cream, bakery and savory products. Cosmetics. Antibacterial . Ayurvedic medicine. Mexican marigold: Lutein . Yellow to orange Baked goods, beverages, breakfast cereals, chewing gum, dairy product Analogs, egg products, fats and oils, sauces, infant and toddler foods. It is also used as antiseptic

23. Green color: 1.Chlorophyll Chlorophylls, the most abundant pigments on earth, are essential for photosynthesis, absorbing light and transducing it into chemical energy Green pigment found in most plants, algae, and cyanobacteria . Chlorophyll a (blue–green) Chlorophyll b (yellow–green) Chlorophyll fat soluble pigment Chlorophyll is converted to chlorophyllins in presence of alkali, which renders it water soluble.

24. minerals OCHRE - Yellow, brown, and red MALACHITE - Green MANGANESE – Black CINNABAR – Red. AZURITE - Blue/dark blue copper LEAD - Red ARAGONITE - Colorless or white mineral LAPIS LAZULI - Blue Tamilarasi and banuchitra , 2021.

25. FRUITS AND VEGETABLES

26. CHEESE Fruit or vegetable Incorporated concentrations Final % Results References Broccoli 3%, 5%, 10%, 15%, 20%, 30%, and 50% 20% All broccoli-cheese powder blended samples contained more total polyphenols as compared to the plain conventional cheese powder. Sharma et al., (2011) Carrot paste 5%, 10%, and 15% 15% The use of carrot paste in cheese improves its quality, antioxidant activity, vitamin A content, and Na/K ratio compared with the control sample. Mohamed et al., (2016) Grape pomace 0.8% and 1.6% 1.6% With addition of grape pomace powders to the cheeses formulation, antioxidant activity and phenolic content in all enriched samples were increased. Marchiani et al., (2016) Tomato extracts 0.5%, 1%, 1.5%, and 2% 2% Sensorial evaluation scores for the yellowness, tomato taste, and hardness of cheese were considerably higher after fortification with tomato extracts (powdered microcapsules). Jeong et al., (2017) Sesame 1%, 3% and 5% 3% The cheese product was found to have the best sensory characteristics at the 3% sesame seed dose compared with the doses of 1% and 5%. Slozhenkina et al., (2019) Spinach powder 0.5%, 1%, 1.5%, and 2% 0.5% and 1% The total solid, protein contents, and acidity of cheese samples were increased considerably with the increasing of spinach nano -powder concentration. El- Sayed , (2020)

27. YOGHURT Fruit or vegetable Incorporated concentrations Final % Results References Strawberry 10%–20% - The addition of strawberry pieces to yogurt can decrease free polyphenols and whey protein contents, possibly constraining its bioaccessibility in yogurt Oliveira et al., (2015) Pineapple peel Powder 1.16% 1.16% Pineapple peel powder addition to yogurt lowered firmness and storage modulus. Syneresis level in probiotic yogurt with pineapple peel powder (1.2% at day 1) was comparable with the prebiotic-inulin And increased during storage. Sah et al., (2016) Carrot juice 0%, 10%, 15%, and 20% 10%–15% Total yogurt carotenoid content was considerably improved with carrot juice enrichment Kiros et al., (2016) Strawberry pulp 15% and 30% 30% The addition 30% of strawberry cryoconcentrates resulted in a product with higher anthocyanins content and antioxidant activity Gasparrini et al., (2017) Pumpkin fiber 0.5%, 1.0%, and 1.5% 1% Pumpkin fiber could improve the physical quality and contributed to textural characteristics of half-fat yogurt. Bakirci et al., (2017) Apple pomace 0.1%, 0.5%, and 1% 1% Apple pomace induced earlier milk gelation and shorter fermentation time, especially at the 1% level Wang, Kristo, et al., (2019) Apple pomace 1%, 2%, and 3% 3% Apple pomace (3%) decreased the syneresis of stirred yogurt by half that of the control. Also, it contributed to total phenolics and dietary fibers to stirred yogurt. Wang et al., (2020)

28. Fruit or vegetable Incorporated concentrations Final % Results References Broccoli 3%, 5%, 10%, 15%, 20%, 30%, and 50% 20% All broccoli-cheese powder blended samples contained more total polyphenols as compared to the plain conventional cheese powder. Sharma et al., (2011) Carrot paste 5%, 10%, and 15% 15% The use of carrot paste in cheese improves its quality, antioxidant activity, vitamin A content, and Na/K ratio compared with the control sample. Mohamed et al., (2016) Grape pomace 0.8% and 1.6% 1.6% With addition of grape pomace powders to the cheeses formulation, antioxidant activity and phenolic content in all enriched samples were increased. Marchiani et al., (2016) Tomato extracts 0.5%, 1%, 1.5%, and 2% 2% Sensorial evaluation scores for the yellowness, tomato taste, and hardness of cheese were considerably higher after fortification with tomato extracts (powdered microcapsules). Jeong et al., (2017) Sesame 1%, 3% and 5% 3% The cheese product was found to have the best sensory characteristics at the 3% sesame seed dose compared with the doses of 1% and 5%. Slozhenkina et al., (2019) Spinach powder 0.5%, 1%, 1.5%, and 2% 0.5% and 1% The total solid, protein contents, and acidity of cheese samples were increased considerably with the increasing of spinach nano -powder concentration. El- Sayed , (2020)

29. ICE CREAM Fruit or vegetable Incorporated concentrations Final % Results References Orange fiber 1.0% and 1.5% 1.0% The orange fiber can be utilized to decrease the fat content (50%) and increase bioactive compounds content, such as dietary fibers and carotenoids. Crizel et al., (2014) Orange by-products fiber 0.74% 0.74% Orange by-products fiber proved to be a promising alternative as a fat replacer in ice cream production. De Moraes Crizel et al., (2013) Carotenoids (extracted from tomato peel) 1%, 2%, 3%, 4%, and 5 2% and 3% SE Ice cream containing 2% and 3% of carotenoids ( lyco -red) had the highest scores for flavor, body and textural properties, melting and color, and the best mix Rizk et al., (2014) Quince seed powder 0%, 0.25%, 0.5%, and 0.75% 0.75% The adding of quince seed powder improved the protein content compared with the control samples due to the high protein content of quince seed powder (35%). The fortification of ice cream with quince seed powder can make an effective way to improve the nutritional and structural properties of ice creams. Atalar , (2018)

30. FDA APPROVAL PIGMENT EEC No YEAR APPROVED USES AND RESTRICTION Annatto extract E160b 1963 Foods generally. Dehydrated beets (beet powder) E162 1967 Foods generally. Butterfly pea flower extract ---- 2021 Fruit and vegetable juice, nutritional beverages, creamers (dairy and non-dairy), ice cream and frozen dairy desserts, hard candy, dairy and non-dairy drinks, fruit preparations in yogurts, and soft candy. Canthaxanthin E161g 1969 Foods generally, 30 mg/lb of solid or semisolid food or per pint of liquid food β- Apo-8'-carotenal E160e 1963 Foods generally , 15 mg/lb solid, 15 mg/pt liquid. β- Carotene E160a 1964 Foods generally. Cochineal extract E120 1967 Foods generally Carmine E120 2009 Food label must use usual name " carmine” chlorophyllin E141 2002 Citrus-based dry beverage mixes  0.2 percent in dry mix; extracted from alfalfa.

31. PIGMENT EEC No YEAR APPROVED USES AND RESTRICTION Grape color extract E163 1981 Non beverage food. Fruit juice ---- 1966 Foods generally. Fruit juice ---- 1995 Dried color additive. Vegetable juice ---- 1966 Foods generally. Vegetable juice ---- 1995 Dried color additive, water infusion. Carrot oil ---- 1967 Foods generally. Paprika E160c 1966 Foods generally. Riboflavin E101 1967 Foods generally. Saffron E164 1966 Foods generally. Spirulina extract ---- 2014 Ice cream and frozen desserts, beverage mixes, yogurts, custards, cottage cheese Tomato lycopene extract; E160 2006 Foods generally. Turmeric E100 1966 Foods generally.

32. Aqueous extraction method – Water soluble pigments Acidic extraction method - Flowers Alkaline extraction method - Phenolic groups, lac insect, annatto seeds, safflower petals Enzymatic extraction method - Bark, root or any hard part of the plants By fermentation - Microbial pigments Turmeric, indigo, annatto seeds Solvent extraction methods - Most of the sources 1) Soxhlet extraction technique - Solid-liquid separation , no filtration , ↑thermal degradation 2) Maceration - Mostly suitable for plant material 3) EXTRACTION BY AGITATION - Degradation of thermo sensitive components Denature heat sensitive pigments Enzymes are costly Bad smell due to microbial action ↑ Yield ↑Quality ↓ Water ↑solvent Extraction methods Chungkrang et al., 2021

33. Ultra sound extraction technique - Polyphenols, carotenoids, anthocyanins (obtained with high purity and safety) . It can deliver high extract yield in a short time using small amounts of solvent (low temperature) Which preserves thermosensitive compounds Pressurized liquid extraction - Curcuminoids from Curcuma longa L., iridoids from Genipa americana L., bixin from Bixa orellana and anthocyanins from Euterpe edulis. 313 to 473 K and pressures between 3.5 and 35 Mpa. Currently, water and ethanol are the most used solvents in PLE processes. Less amount of solvent is used and a shorter process time is required . Not suitable for extraction of thermosensitive colorants. HHP - It uses pressure between 100 and 1,000 M pa at room temperature @ 3 and 10 min. Extraction of genipin from genipap fruit, extraction of phenolic compounds from watercress, and extraction of anthocyanins from apple. HHP can be used to extract bioactive compounds from plants. Currently, used in most of the dairy products. Reduced lipids degradation, does not generate waste by-products. High cost.

34. Supercritical fluid extraction A supercritical fluid has viscosity similar to a gas, density similar to a liquid and diffusivity between gas and liquid, which favors the extraction of intracellular compounds More recently, SFE has stood out as a potential technique to recover nutritionally valuable compounds, mainly thermo sensitive ones , from solid or liquid matrices Supercritical carbon dioxide (SC-CO2) is the solvent most used in SFE . The use of SC-CO2 is recommended to recover carotenoids and chlorophylls. This solvent is not suitable for the extraction of betalains and anthocyanins .

35. Conclusion and future prospects: Objective of adding colour to foods is to make them appealing and it consumer to buy a product. Synthetic food colours cause carcinogenicity, toxicity, allergenicity, and other effects on human health. Plant based pigments are mostly used in dairy products compare with animal and microbial sources. Solvent extraction techniques are frequently employed to extract natural pigments (DME can extract pigments or lipids from microalgae without any preliminary drying and cell disruption steps) The development of low-cost, feasible extraction technology for the production of natural colours and its application in the dairy industry is a challenging and current necessity.

36. Some flowers used for natural dying and their colour observed Sarkar et al., 2023.

37. THANK YOU for your valuable time….

38. References Beyer, P., Al- Babili , S., Ye, X., Lucca, P., Schaub , P., Welsch , R., & Potrykus , I. (2002). Golden rice: introducing the β- carotene biosynthesis pathway into rice endosperm by genetic engineering to defeat vitamin A deficiency.  The Journal of nutrition ,  132 (3): 506S-510S. Chaitanya Lakshmi , G. (2014). Food coloring: the natural way.  Res J Chem Sci ,  2231 (8): 606X. Chaitanya Lakshmi , G. (2014). Food coloring: the natural way.  Research Journal of Chemical Sciences ,  2231 (8):606X. Druding SC. Dye History from 2600 BC to the 20 th Century. Seminar presented in Seattle, Washington at Convergence a bi-annual gathering of weavers, dyers and spinners; c1982. Dufossé , L. (2018). “Microbial pigments from bacteria, yeasts, fungi, and microalgae for the food and feed industries,” in Handbook of Food Bioengineering: Global data consumer survey, (2022). Natural and Artificial Flavoring Agents and Food Dyes, Vol. 7, eds A. M. Grumezescu and A. M. Holban (Amsterdam: Academic Press), 113–132.

39. Sarkar , D., Khan, A. M., Chakraborty , O., Sarkar , I., & Maitra , S. (2023). Natural dye: Antiquity to future perspective.  The Pharma Innovation Journal ,  12 (6) :1536-1544. Salehi , F. (2021). Quality, physicochemical, and textural properties of dairy products containing fruits and vegetables: A review.  Food Science & Nutrition , 9 (8): 4666-4686. Rana , B., Bhattacharyya, M., Patni , B., Arya , M., & Joshi, G. K. (2021). The realm of microbial pigments in the food color market.  Frontiers in Sustainable Food Systems ,  5 : 603892. Tamilarasi , A., & Banuchitra , M. (2021). Classification and types of natural dyes: a brief review.  International Journal of Creative Research Thoughts (IJCRT) ,  9 (11): 527-532. Sen , T., Barrow, C. J., and Deshmukh , S. K. (2019). Microbial pigments in the food industry-challenges and the way forward. Frontiers in Nutrition . 6: 1–14. U.S Food and Drug administration, (2023).