Exposure assessment to synthetic food colours of a selected population in Hyderabad, India

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1 Food Additives and Contaminants, Vol. 21, No. 5 (May 2004), pp Exposure assessment to synthetic food colours of a selected population in Hyderabad, India Pratima Rao{*, R. V. Bhat{, R. V. Sudershan{, T. P. Krishna{ and N. Naidu{ { Food & Drug Toxicology Research Centre, National Institute of Nutrition, Jamai-Osmania (P.O.), Hyderabad , India { National Institute of Nutrition, Jamai-Osmania (P.O.), Hyderabad , India (Received 27 June 2003; revised 16 January 2004; accepted 27 January 2004) An exposure assessment of synthetic food colours was carried out among 1 5- and 6 18-year-old individuals by the food frequency method. Children had an intake of solid food consumption in the range g day 1 and liquid food consumption in the range ml day 1 with added colours. Among the eight permitted colours in India, six were consumed by the subjects of the study. The intakes of some subjects exceeded the acceptable daily intake for colours such as tartrazine, sunset yellow and erythrosine, which is 7.5, 2.5 and 0.1 mg kg 1 body weight, respectively. Therefore, a uniform permissible limit of 100 mg kg 1 prescribed under the Prevention of Food Adulteration Act in India for all foods is not justified. The limits need to be revised according to the Codex Alimentarius Commission, which permits different maximum levels of additives to various food categories based on both the extent of consumption and the technological justification for its use. Keywords: exposure assessment, food colours, tartrazine, sunset yellow, erythrosine, acceptable daily intake (ADI) Introduction The potential for adverse health effects through exposure to naturally occurring or synthetic agents in * To whom correspondence should be addressed. jrpratima@ yahoo.com foods is evaluated by risk analysis. Among the synthetic agents, food colours are of major concern in India. To evaluate the safety of food colours, the Joint FAO/WHO Expert Committee of Food Additives (JECFA) has set an acceptable daily intake (ADI) for each of the permitted colours based on toxicological studies on experimental animals and data from human clinical studies. The ADI has been defined as the amount of a substance that can be consumed everyday throughout the lifetime of an individual without any appreciable adverse health effects (JECFA 1996). The ADI for permitted colours varies from 0.1 mg kg 1 body weight for erythrosine to 25 mg kg 1 body weight for fast green FCF. It becomes extremely important to monitor the total daily intake of all food colours since the ADI of none of the colours should be exceeded (World Health Organization 1991). The ILSI Europe Acceptable Daily Intake Task Force suggested that special ADIs should be established for infants and children as the use of food colours represents a higher risk for infants and children than for adults. Children, in general, are more susceptible to chemical insults and the patterns of consumption are different from adults (Larsen and Pascal 1998). The use of permitted colours has raised concerns that they might be used in excess of the statutory limit (100 mg kg 1 ) or in foods in which they were not permitted (Bhat and Mathur 1998, Padmaja et al. 2001, Pratima Rao and Bhat 2003). The safety of repeated exposure to permitted synthetic colours has been questioned (Lockey 1977). The ADI of erythrosine was reduced from 2.5 to 0.1 mg kg 1 body weight as it produced toxic effects on thyroid function in short-term studies in rats (Larsen 1991). Permitted colours like ponceau 4R, tartrazine and sunset yellow have provoked allergic reactions in several individuals, even at low levels of intake (Lockey 1977). The allergic responses vary from urticaria to dermatitis, angio-oedema and exacerbation of the asthmatic patients (World Health Organization 1991). It has also been reported that the consumption of a particular Food Additives and Contaminants ISSN X print/issn online # 2004 Taylor & Francis Ltd DOI: /

2 416 P. Rao et al. brand of aniseed having very high levels of ponceau 4R exhibited symptoms of glossitis of the tongue in children (National Institute of Nutrition 1994). Tartrazine was associated with irritability, restlessness and sleep disturbance in atopic or hypertensive children aged between 2 and 14 years (Rowe and Rowe 1994). In the USA and Italy, it was observed that people are more exposed to indigotine, tartrazine and sunset yellow, while in Switzerland the population was exposed to colours like ponceau 4R, azorubine and amaranth (Toldeo et al. 1992). The Prevention of Food Adulteration Act in India prescribes a level of 100 mg kg 1 for tartrazine, sunset yellow, ponceau 4R, carmoisine, erythrosine, brilliant blue, fast green FCF and indigo carmine to be added to specified food items and a level of 200 mg kg 1 in foods in general such as canned foods, jams and jellies. There is a need to fix the levels based on the intake of specified food colours through different foods and the ADI for that colour. Thus, it becomes essential to know the intake of food colours to ensure that with increasing authorized use, the ADI is not exceeded by the population. Therefore, an attempt was made at an exposure assessment of synthetic food colours in a selected population of preschool and school-going children in urban and rural areas of Hyderabad, India. Materials and methods Preschool (1 5 years) and school-going (6 18 years) children in urban Hyderabad from three socioeconomic income groups such as high, middle and low income during summer and winter were selected for study. From the selected 525 households, there were 175 households each from high-, middle- and low-income groups. Of the 175 households in each category, 100 from each income group were selected for winter and 75 households from each income group were selected randomly. Since it was a household survey, the homemaker of the household was interviewed for the intake of foods with added colours by all the members of the family. Three hundred households from three districts, Nalgonda, Mahboobnagar and Ranga Reddy district, A.P., were selected for study (table 1). Thus, 662 individuals in urban areas and 358 in rural areas were surveyed. The diet survey was carried out by a food Table 1. Profile of individuals surveyed in urban and rural areas. Age group (years) High income Middle income Low income Rural Total frequency method as foods with added colours are of habitual consumption. Information on the habitual frequency of consumption of foods with added colours was elicited using a coloured food atlas that was computed during a market survey to enable the respondents to recall the foods they consume. The portion sizes, i.e. quantities of foods with added colours consumed, were recorded using standardized stainless steel cups and vessels. To quantify the amount of beverage intake, standard measuring glasses of 60, 120, 240 and 500-ml capacities were used. The foods with added colours consumed by the respondents in both urban and rural areas were procured from local markets, vendors, supermarkets, sweetmeat stalls, etc. and were analysed for the type and level of colours. Qualitative analysis was conducted using the wool dyeing and column chromatography techniques for extraction of colours, followed by identification through ascending paper chromatography in standard solvent systems such as 1% ammonia, 2% sodium chloride in 50% ethanol and phenol (80 g) in 20 ml water (Toteja et al. 1990). Quantitative analysis was carried out by measuring the optical density of the coloured solutions using a spectrophotometer (Ranganna 1986). Replicate analysis of the same foods was undertaken as a part of an interlaboratory quality assurance programme carried out by three established research centres in India. Spiked food samples were exchanged between the centres and the qualitative and quantitative determination of synthetic colours in these foods was undertaken. The results were comparable and, hence, for further analysis the same methods were used (table 2). For foods containing more than one synthetic colour, each colour was eluted separately from the paper chromatography and their optical densities were measured by spectrophotometry. The mean colour intakes per day for each individual were calculated from the analytically determined concentrations of colours present in the ingested foods.

3 Exposure assessment to synthetic food colours of a selected population in Hyderabad 417 To evaluate the potential risk associated with the consumption of synthetic colours by the population surveyed, a comparison was made between the mean colour intakes observed in this survey and the ADIs recommended by JECFA. The mean colour intake per day by each individual was calculated from the concentration of colours in foods in order to determine the ADI of the colours ingested. The ADI was calculated using the formula: Colour ingested per day/body weight of individual. The ADI is the acceptable daily intake (that denotes the amount of a substance that can be consumed daily throughout the lifetime of an individual without any appreciable health effects) and has been used to evaluate the risk associated by consumption of synthetic food colours by the selected population. Since the weights of children were not taken at the time of survey, the standard National Centre for Health Statistics (NCHS) weight for height/age charts were used to derive the weights of the children. Table 2. Sample no. Detection of colours in spiked sugar samples. Colours detected Amount spiked Centre I Centre II Centre III A tartrazine B sunset yellow C tartrazine sunset yellow A tartrazine B sunset yellow C tartrazine sunset yellow A tartrazine B sunset yellow C tartrazine sunset yellow Results The intake of the foods with added colours intake among 1 5- and 6 18-year-old groups is shown in table 3. Both groups consumed solid foods with added colours in the range g and liquids with added colours in the range ml. There was not much difference in the mean consumption of the individual foods with added colours in both groups. However, within each group, a wide variation was observed in the quantity of the coloured foods consumed. The beverages consumed by both groups included fruit drinks, squashes and sherbets. The 6 18-year-old group in addition consumed carbonated beverages. The type of sweetmeats (the term in India denotes food preparations made from cereal or pulse or milk as the basic ingredients along with sugar) consumed by both age groups varied. The 1 5-year-old children consumed sweetmeats, i.e. mysore pak (pulse based fudge) and halwas, while the 6 18-year-old group consumed sweetmeats like laddu, jilebi (deep fried delicacy of fermented batter in spiral shapes sweetened in sugar syrup) and burfi (a sweetmeat preparation of North India made from evaporated milk, sugar, water and nuts). Confectioneries in the form of peppermints, sugar toys and lollipops were consumed by the 1 5-year-old children while the 6 18-year-old group consumed chewing gums, peppermints, mouth fresheners, fruit jelly, sugar candies and sugar toys. Miscellaneous products like soups, jams, savouries, custard, ice candies and starters such as manchuria were consumed by both groups; in addition, the 6 18-year-old group consumed sauces, crushed ice, aniseed, etc. Subjects from rural areas were observed to have a lower consumption of foods with added colours, i.e. solid foods in the range g and liquids in the range ml. The varieties of foods consumed were limited such Table 3. Coloured food consumption pattern among individuals of 1 5 and 6 18 years of age. Age (years) Beverages (ml) Sweetmeats (g) Confectionery (g) Bakery (g) Miscellaneous (g) 1 5 number of children mean SD range number of children mean SD range

4 418 P. Rao et al. as beverages like synthetic syrups packed in plastic sachets or plastic containers, sweetmeats like laddus, mysore pak and jilebi, confectionery (denotes preparations made from sugar as the basic ingredient that may be hard or soft boiled) like lollipops, peppermints, sugar toys, bakery items like rusk and biscuits and miscellaneous foods like savouries, and crushed ice. The foods consumed by the subjects were analysed for the type and level of colours (table 4). Of the eight permitted colours prescribed by the Prevention of Food Adulteration, India, six were detected, while the two colours such as fast green FCF and indigo carmine were not detected in any of the foods analysed. The prescribed levels of colours like tartrazine were exceeded in a majority of foods like sweetmeats, the concentrations being mg kg 1 in sweetmeats and 9450 mgml 1 in beverages, while among rural hard-boiled confectionery foods like sugar toys, the concentration of ponceau 4R was maximum (4001 mg kg 1 ) and in mouth fresheners, brilliant blue was detected in high concentrations such as 4673 mg kg 1 and mg kg 1 in savouries. The mean total colour and mean individual colour intake among 1 5- and 6 18-year-old individuals showed that the mean total colour intake was about 17.2 mg and was the same in both groups. Among the eight permitted colours in India, six were consumed by the subjects of the study. Tartrazine and sunset yellow were predominantly consumed by both groups through consumption of sherbets containing high concentrations of tartrazine (9450 mg l 1 ) along with ice candies and sweets such as jilebi, pedas, halwas and burfis. Carmoisine and ponceau 4R were ingested through consumption of sherbets, sweetmeats (pedas, burfis) and jams by both groups. Brilliant blue was consumed through green-coloured sweetmeats, confectionery, sherbets and savouries in combination with tartrazine. The results indicate that the colour intakes of only 3% of the 1 5-year-old group and by 2% of the 6 18-year-old group was above the ADI. This represented particular instances on particular days when the children had ingested foods containing high concentrations of synthetic colours. The tartrazine intake of three children with an intake of mg of the colour from the intake of beverages and savouries and the sunset yellow intake of four children who ingested mg of the colour through sweetmeats, beverages and savouries exceeded the ADI on those days of consumption. Among the 6 18-year-old group, the ADI of three colours was exceeded for 13 individuals. The instances when the ADI was exceeded were when four children ingested mg tartrazine through beverages, savouries and miscellaneous foods like custard and soups, and among seven children who ingested mg sunset yellow through sweetmeats, beverages and miscellaneous foods like chegodi (deep fried savoury made from rice flour), sev (extruded product made from pulse flour), boondi (deep-fried savoury made from pulse flour) and finger fries, biryani (a rice preparation), crushed ice and sauce. The intake of erythrosine exceeded the ADI among two children, which was 4.9 mg through the consumption of confectionery and pastries (table 5). Table 4. Concentration of permitted food colours in various foods from urban and rural areas. Food items Region Range of permitted colours (mgg 1 or mgml 1 ) Tartrazine Sunset yellow Ponceau 4R Carmoisine Brilliant blue Erythrosine Sweetmeats urban rural Breakfast accompaniments urban rural Beverages urban rural Confectioneries urban rural Bakery products urban rural Miscellaneous urban rural

5 Exposure assessment to synthetic food colours of a selected population in Hyderabad 419 Table 5. Mean permitted colour intake among individuals of 1 5 and 6 18 years of age, and profile of acceptable daily intake. Age groups (years) Permitted colours Range (mg) Mean colour intake (mg) Actual intake of colour (mg kg 1 body wt) Acceptable daily intake (%) 1 5 tartrazine sunset yellow carmoisine ponceau 4r brilliant blue erythrosine total tartrazine sunset yellow carmoisine ponceau 4r brilliant blue erythrosine total Body weight of 1 5 years, 19.7 kg; 6 18 years, 21.7 kg (NCHS Standards). 18% 14% 21% 18% 10% 10% 42% 16% Figure 1. Level of exposure among 1 5 yrs children during winter. 17% 17% 18% 22% 26% Figure 2. Level of exposure among 1 5 yrs children during summer. 23% 28% Figure 3. Level of exposure among 6 18 yrs children during winter. individuals had maximum exposure from bakery items, i.e. rusk, cakes, pastries and biscuits, followed by confectionery (23%) such as lollipops, peppermints, bubblegums and mouth fresheners (figure 3). However, during summer, the subjects of 6 18-yearold group had maximum exposure from confectionery, i.e. sugar candies, sugar toys, lollipops and chewing gums, followed by beverages (24%) such as sherbets and fruit drinks (figure 4). The extent of exposure of individuals to added colours from various foods during summer and winter was studied. During winter, the maximum colour intake (42%) was from confectionery followed by beverages (18%) (figure 1). In summer, the maximum colour intake was from beverages (26%) in the form of carbonated beverages, fruit drinks and sherbets (figure 2). During winter, the 6 18-year-old Discussion The total colour intake in India per person was calculated as 17.2 mg, which is less than in the USA, where it was 77.1 mg (Johnson and Litchtenberger 1984). The lower intake of colours in India is due

6 420 P. Rao et al. 19% 20% and the maximum level ranges from 3.1 to 500 mg kg 1, while erythrosine has been permitted in 44 food categories in the range mg kg 1. 27% 10% 24% Acknowledgements Sweetmeat Beverages Bakery Confectionary Miscellaneous Figure 4. Level of exposure among 6 18 yrs children during summer. The authors thank the Director, National Institute of Nutrition (NIN), for the keen interest shown in the study. The financial support provided by the Department of Food Processing, Government of India, New Delhi, is gratefully acknowledged. to the lower consumption of processed foods. Tartrazine and sunset yellow are the colours most commonly used in India as found in the present study. In countries like Brazil (Toledo et al. 1992) and the USA (Johnson and Litchtenberger 1984), these were also the most commonly used colours. However, the use of other colours varied between countries. Fast green FCF and indigo carmine were not consumed by subjects of the study. This would mean that the food industry does not find it necessary to use these colours. Hence, it may be worthwhile to consider the withdrawal or clearance of these two colours in consultation with industry. The ADIs of tartrazine, sunset yellow and erythrosine were exceeded in a few children on particular days of consumption, i.e. by 104, 284 and 200%, respectively. The ADI of tartrazine and sunset yellow was exceeded by the consumption of foods having high concentrations of colours, while the ADI of erythrosine was exceeded when the intake of processed foods was normal. For example, it was observed that by the intake of only one pastry weighing 200 g, the intake of erythrosine was 4.8 mg, which exceeded the ADI by 200%. The ADI of permitted colours varies from 0.1 to 25 mg kg 1 body weight. Therefore, the fixing of a uniform limit of 100 mg kg 1 for all foods as is currently done under the Prevention of Food Adulteration Act is not justified. According to the Codex Alimentarius Commission (1999), maximum levels of additives to various food categories should be based on both the extent of consumption and the technological justification for its use. For example, tartrazine has been permitted in 80 food categories References Bhat, R. V., and Mathur, P., 1998, Changing scenario of food colours in India. Current Science, 74, Codex Alimentarius Commission, 1999, Discussion paper on the use of colours in foods. Joint FAO/WHO Food Standards Programme, Codex Committee on Food Additives and Contaminants CX/FAC 00/9. Agenda item 8, December 1999 (Codex Alimentarius Commission). Rome. Johnson, R. K., and Litchtenberger, F. J., 1984, Synthetic certified food colours of the USA. Developments in Food Colours 2, edited by J. Walford (London: Elsevier), p Joint Expert Committee on Food Additives, 1996, Summary of Evaluations Performed by the Joint FAO/WHO Expert Committee on Food Additives, FAO/IPCS/WHO (Geneva: WHO). Larsen, J. C., 1991, Erythrosine: toxicological evaluation of certain food additives and contaminants. 37th Meeting of JECFA/ WHO Food Additive Series, 28, pp Larsen, J. C., and Pascal, G., 1998, Workshop on the applicability of the ADI to infants and children: consensus summary. Food Additives and Contaminants, 15, Lockey, S. D., 1977, Hypersensitivity to tartrazine (FD & C Yellow #5) and other dyes and additives present in food and pharmaceutical products. Annals of Allergy, 38, National Institute of Nutrition, 1994, Studies on Newer Adulterants and Contaminants. Annual Report (Hyderabad: Indian Council of Medical Research). Padmaja, R. J., Pratima Rao, Bhat, R. V., and Naidu, A. N., 2003, Type, extent and use of colours in ready-to-eat (RTE) foods prepared in the unorganized sector Hyderabad, India a case study. International Journal of Food Science and Technology in press, 38, 1 7. Pratima Rao, and Bhat, R. V., 2003, A comparative study on the synthetic food colours usage in foods procured from urban and rural areas of Hyderabad. Nutrition and Food Science, 33, Ranganna, S., 1986, Handbook of Analysis and Quality Control for Fruits and Vegetable Products (New Delhi: Tata McGraw-Hill). Rowe, K. S., and Rowe, K. J., 1994, Synthetic food colouring and behaviour: a dose response effect in a double blind, placebo controlled repeated measures study. Journal of Paediatrics, 125,

7 Exposure assessment to synthetic food colours of a selected population in Hyderabad 421 Roy, A. K., and Chakraborti, J., 1991, Added colours in foodstuffs from Calcutta markets. Science and Culture, 57, Toledo, M. C. F., Guerchon, M. S., and Ragazzi, S., 1992, Potential weekly intake of artificial food colours by 3 14 year old children in Brazil. Food Additives and Contaminants, 9, Toteja, G. S., Mukherjee, A., Mittal, R., and Saxena, B. N., 1990, ICMR Manual Methods of Analysis for Adulterants and Contaminants in Foods (New Delhi: Indian Council of Medical Research). World Health Organization, 1991, Toxicological Evaluation of Certain Food Additives and Contaminants. Food Additives Series 28 (Geneva: WHO).

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