Iii

5 ng/ml of caramel IV 50, 150 or 500 mg/kg bw/day/109 wk following in utero exposure 50, 150, or 500 mg/kg bw/day/three generation 0, 250, 500, and 1000

mg/kg bw/day/90 days 320, 800, 2000, 5000 mg/kg bw/day, oral or intraperitoneally, evaluations up to the seventh day

Results

No mutagenic effect

No mutagenic effect

Neither mutagenic nor chromosomal damage observed; thus, it is concluded that caramelization procedures do not produce detectable levels of mutagens

No clastogenic

No clastogenic effect

No clastogenic effect No carcinogenic activity

No adverse effects observed

No toxicity exhibited but only reduced growth rates at the two highest doses Aspartate and alanine aminotransferase show dose-dependent increments; maximum enzymatic activity is reached at 24 h followed by a decrement; diarrhea is observed after 3-4 h with feces dyed blue at doses more than 800 mg/kg; intraperitoneal doses do not produce increments in the enzymatic activity but up to a dose of 2000 mg/kg; hemorrhagic lesions and mortality are observed at concentrations higher than 2000 mg/kg; thus, gardenia color induces hepatotoxicity and is mainly associated with its iridoid colorant, geniopside

TABLE 4.5 (continued)

Studies on Toxicity and Safety of Colorants Exempt from Certification

Color

Gardenia blue colorants

Riboflavin

Model

Mice

Mutagenicity Umu test, SOS Chormotest, and Ames Salmonella assay

Conditions

0.15, 0.30, and 0.60% dietary from 5 weeks of age in F0 generation to 9 weeks of age in the F1 generation; mating at 9 weeks of age

0-100 mg/mL, with or without activation (S9 or cecal extract)

Results

No differences in food consumption, growth, or survival; a significant increase in the mean value of the left lung weights in males at 5% dose; non-neoplastic lesions appear in the control and treated groups but without differences between them; tumors in different organs but not associated with color feeding; it is concluded that gardenia color does not exert any carcinogenic effect

In the F0 generation, increased food consumption in a dose-dependent manner; no effects on body weight, gestation or lactation periods, exploratory behavior

In the F1 generation, no effects in body weight and survival, but behavioral development parameters show tendencies to be depressed (surface righting, cliff avoidance, and swimming); olfactory orientation is accelerated; it is concluded that doses permitted in food human consumption are below the evaluated levels and lac dye produces no adverse effects in humans

Lumifavin, its activated metabolite, has also been analyzed; riboflavin does not show mutagenicity but activated lumifavin shows a significant effect in all the tests; it is suggested that the activated product acts as an intercalating DNA product

Monascus pigments

Mutagenicity Salmonella, hepatocyte and microsome assays

0, 50, 100, 500, 1000, and 5000 ^g/plate, comparison with citrinin

In all Monascus extracts citrinin is identified at up to 1.8 ng/g extract and citrinin is a mycotoxin; no mutagenicity has been detected in the Salmonella microsome assay with or without activation; in the Salmonella hepatocyte assay, mutagenicity is dose dependent and this is related with the effect of citrinin; thus, it has been suggested that Monascus mutagenicity is mediated by a complex activation processing in hepatocytes

TABLE 4.5 (continued)

Studies on Toxicity and Safety of Colorants Exempt from Certification

TABLE 4.5 (continued)

Studies on Toxicity and Safety of Colorants Exempt from Certification

Color

Model

Conditions

Results

Turmeric

Rat

500 mg/kg bw/1 year or

No significant effect

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