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Abstract
The snack product marketing has been grown steadily in the last few years. It is reported that sales of corn-based snacks reached 1.6$ billion in 1985, up from 1.1$ billion in 1980. Some prepared foods and snacks which are consumed by children obtain food additives. Coloring agents, antioxidants, preservatives and nutritional supplements are added to food additives have many benefits, however they also have negativeaspects, it has been argued that many food additives, particularly flavorings and colors, offer little if any practical value. One of the most common negative claims about food additives is that they can provoke exacerbate a variety of behavioral problems including depression, blurred vision, dizziness, insomnia, nervousness, migraines, and hyperactivity. Unfortunately, there are few published data concerning the quality control of processed foods including food additives in Egypt in particular infant formula. Therefore, the present study aims to study the biological evaluation of some food additives distributed in Egyptian local markets in vitro. To achieve like of these aims, a total of thirty samples of different types of snakes i.e. potato chips, loleta, karate, (10 samples per each) were collected at random from the supermarkets of Shibin El-Kom City, Minufiya Governorate, Egypt. All samples were purchased within about one month, transported to the laboratory, ground or homogenized before they were freeze-dried to ensure equal moisture content then used for food additives extraction. The all resulted extracts were used for the all cytotoxiciy and genotoxicity assays. The obtained results could be summarized as follow: 1. Studying the potential cytotoxic effects of tested food additives extracts towards liver homogenate of karmoot fish. Catfish liver cells homogenate were used as an experimental tool for studying the toxic, biochemical, and immunological effects of five synthetic and natural food additives extracted from snacks distributed in Egyptian local markets. For cytotoxic effects, NR, MTT, and CV assays which determined the lysosomes activity, mitochondrial activity, and cell membrane integrity of liver cells, the absorbance measurements of assays (as % of control) were 14.55 – 109.21, 31.23 – 99.88, 25.23 – 113.42, 54.34 – 124.22 and 78.89 – 126.22 ( for NR); 19.48 – 101.00, 33.78 – 100.09, 27.56 – 114.43, 52.46 – 125.72 and 88.89 – 123.77 ( for MTT); and 22.20 – 116.70, 35.67 – 99.09, 27.09 – 110.34, 68.39 – 130.90 and 90.89 – 126.67 (for CV) for the previous extracts, respectively. The highest adverse cytotoxic effects were recorded for synthetic additives i.e. Karate snacks followed by loleta and potato chips, respectively. A very little effects induced by natural extracts (curcumin and beet roods). Also, it could be easily concluded that, NR assay is more sensitive to all tested under this investigation than others assays. 2. Studying the adverse biochemical effects of tested food additives extracts towards liver homogenate of karmoot fish. The influence of five synthetic and natural food additives extracted from snacks distributed in Egyptian local markets on the growth assay which performing by protein determination (GA) and protease activity (PA) of liver cells homogenate were determined in a trial to make a comparison between biochemical and cytotoxicity assays. The absorbance measurements of assays (as % of control) were 30.70 – 102.35, 35.99 – 98.90, 29.67 – 111.23, 79.87 – 126.12 and 90.88 – 119.00 ( for GA); and 30.78 – 98.21 – 40.96 – 100.06, 33.78 – 121.32, 86.89 – 132.29 and 93.89 – 130.29 (for PA) for the previous extracts, respectively. The highest adverse cytotoxic effects were recorded for synthetic additives i.e. Karate snacks followed by loleta and potato chips, respectively. A very little effects induced by natural extracts (curcumin and beet roods). The present data demonstrated that the dose-response curves with the NR, MTT and CV assays corresponded well to those with GA and PA assays. 3. Studying the initial and midpoint toxicity of the tested food additives extracts towards liver homogenate of karmoot fish. To analyze the cytotoxicity data it was necessary to determine the exposure concentration to tested food additive extracts causing initial toxicity (NR, MTT , CV , GA , and PA values) and those causing 90 90 90 90 90midpoint toxicity (NR, MTT , CV , GA and PA values). Such data 50 50 50 50 50were necessary to distinguish and/or for comparison amongst the all tested extracts. For example, the midpoint cytotoxicity value for karate snacks extract was recorded lowest values for NR, MTT, CV, GA and PA followed by loleta and potato chips extracts. The natural extracts in particular beet roods no recorded values i.e. out of the curve dimensional for the almost cytotoxic and biochemical assays. According to these data, the sequence of tested extracts for the different cytotoxicity assays were karate snacks> loleta> potato chips> curcumin while beet roods recorded no effects. 4. The toxicity and mutagenic potential of synthetic and natural food additives extracted from snacks distributed in Egyptian local markets in human lymphocyte cultures: Viability of human lymphocyte: The viability of human lymphocyte exposure to synthetic and natural food additives
extracted from snacks distributed in Egyptian local markets. The 7 -6 -5 -4 -3 -2 -1tested dilutions (10 , 10 , 10 , 10 , 10 , 10 , 10 ) of synthetic food additives decreased the human lymphocyte viability which were 40.67, 37.50, 35.70, 31.91, 30.20, 28.04 and 24.77% (for karate snack), 46.98, 43.83, 39.54, 36.77, 33.67, 34.54 and 31.87% (for potato chips); and 43.51, 39.45, 37.44, 34.90, 30.20, 30.00 and 28.90% (for loleta). A very little effects induced by natural extracts (curcumin and beet roods) especially at the high concentrations. The present data demonstrated that the dose-response curves with the NR, MTT and CV assays corresponded well to those with GA assay. DNA damage detection by comet assay: The tested dilutions -7 -6 -5 -4 -3 -2 -1(10 , 10 , 10 , 10 , 10 , 10 , 10 ) of synthetic and natural food additives increased the percentage of total damaged spots (plate III) with values of 97.89, 96.15, 94.88, 94.44, 90.02, 89.99 and 87.04% (for karate snack), 98.90, 9.99, 97.87, 96.99, 95.03, 95.55 and 94.67% (for potato chips); 97.99, 96.15, 96.00, 95.66, 93.00, 93.33 and 92.93% (for loleta), respectively when compared with control samples. A very little effects induced by natural extracts (curcumin and beet roods) especially at the high concentrations. These values were found to be highly significant p<0.001) and dose dependent.