Abstract
4-Dimethylaminoazobenzene (DAB, methyl yellow, or butter yellow), a human carcinogen, has been banned for use in foods since 1988. In 2014, DAB adulteration in Tofu occurred in Taiwan. We hypothesize that DAB can form [DAB•SBP]adduct adduct with soybean protein (SBP) which could damage Gastro-Duodenal-Hepatic axis. Sprague-Dawley rats gavage fed [DAB•SBP]adduct adduct revealed severely reduced body weight and damaged duodenum, liver, hepatic mitochondria, and spleen. Hepatic levels of glutathione and ATP were severely reduced. Serum GOT and GPT were substantially elevated. Analysis by the adsorption isotherm clearly revealed DAB formed very stable [DAB•SBP]adduct adduct at 1:1 molar ration (Phase A). The equilibrium constant of this colloidal adduct [DAB•SBP]adduct was KeqA = ∝, behaving as the most stable and toxic species. At higher protein concentration (Phase C) it formed conjugate [DAB×SBPgross]conjugate, with KeqC = 3.23×10−2 mg/mL, implicating a moderately strong adsorption. The in vitro pepsin digestibility test showed apparently reduced digestibility by 27% (by Ninhydrin assay) or 8% (by Bradford assay). Conclusively, this is the first report indicating that [DAB•SBP]adduct potentially is capable to damage the Gastro-Duodenal-Hepatic axis.
Highlights
Azo dyes are compounds characterized by their vivid colors and provide excellent coloring properties
Azo dyes account for approximately 60–70% of all dyes used in food, textile and leather industries[1]. 4-Dimethylaminoazobenzene (DAB, methyl yellow, or butter yellow), and o-amino-azotoluene are typical aminoazo dyes
To 5 mL of soybean protein (SBP) solutions at different concentrations (0 to 220 mg SBP/mL) 300 μL of experimental DAB solution (1.1265 mg/mL) was added, agitated at 25 ̊C for 30 min and centrifuged for 3 h at 12,000×g, the supernatant was read at 430 nm
Summary
Azo dyes are compounds characterized by their vivid colors and provide excellent coloring properties. 3’Me-DAB forms t-RNA- azo dye complex in liver of rats, which is considered as the major target for this azo carcinogen[9]. The levels of DAB in these products ranged from 2.1 to 63.6 parts per billion (ppb) In reality, this DAB scandal was first set off by Hong Kong’s Center for Food Safety who discovered the prohibited dye in prepackaged pepper-flavored hard Tofu curds manufactured by Te-Chang Food Co. It was hypothesized that the adulteration of DAB into Tofu in vitro could form dye–protein adducts which may retard the digestibility of protein and simultaneously damage the gastroenteric system when ingested per os and it was supposed that 2–3 week-chronic ingestion should be able to induce the same event in the animal model. We conducted the in vitro pepsin digestibility study as well as the in vivo animal experiment using the DAB-soybean protein model
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