Simultaneous Determination And Cytotoxicity Of Several Synthetic Colorants

Synthetic colorants have been widely used in food. And the rapiddetermination and Cell toxicity of synthetic colorants is warranted. Thus, Method forsimultaneous determination of21kinds of synthetic colorants in foods by liquidchromatography were developed. On this basis, it is easy to determine7kinds ofsynthetic colorants in foods by liquid chromatography tandem mass spectrometry. Fortoxicity evaluations, cellular morphology, mitochondrial function (MTT assay) wereassessed under control and exposed conditions. The results are as follows：1. An analytical method based on HPLC has been developed for simultaneousdetermination of Tartrazine, Amaranth, Indigo, Carmine, Brilliant Black BN, SunsetYellow, Allura Red, Acid Red2G, Azorubin, Acid Red26, Lissamine Green B,Brilliant Blue, Acid Orange I, Erythrosin B, Acid Red52, Orange Ⅱsodium salt,Patent Blue V, Acid Yellow36, Auramine, Rhodamine B and Chrysoidine G in foods.HPLC was carried out on the C18column (4.6mm×250mm,5μL), by means of adiode array detector, with acetonitrile/0.05mol/L ammonium acetate gradient elutionand at the flow rate of1.0mL/min, at the column temperature of35°C. The detectionwavelength was240,410,510,627nm. There were good linearity in all pigmentbetween2~40μg/mL. Their recoveries were above75%and the limits of detectionwere2~10μg/mL.2. A comprehensive analytical method based on liquid chromatography tandemmass spectrometry has been developed for simultaneous determination of Acid red2G,Azorubin, Acid Red26, Acid Red52, Patent Blue V, Lissamine Green B andQuinoline Yellow in foods. The samples were extracted, purified by small polyamidecolumn and then qualitative and quantitative analyses were carried in negative modeunder the multiple reaction monitoring (MRM) mode after the chromatographicseparation on XDB-C18(3.5μm,4.6×100mm) column with acetonitrile-ammoniumacetate gradient elution. Under the optimum conditions, the limits of detection (LODs)for Acid red2G, Azorubin, Acid Red26, Acid Red52, Patent Blue V, Lissamine Green B and Quinoline Yellow were0.25-30ng/mL. There were good linearitybetween0.5-1000ng/mL. The mean recovery was over85%and the relative standarddeviation (RSD) was less than10%.3. In Vitro Cytotoxicity of synthetic colorants in Human Liver Cell Line HL-7702：The aim of the study is to assess the toxicity of Acid Red2G, Auramine, ChrysoidineG, Patent Blue V, Lissamine Green B and Quinoline Yellow in liver cells. For toxicityevaluations, cellular morphology, mitochondrial function (MTT assay) were assessedunder control.The microscopic studies demonstrated that each group which compared with thecontrol group had different effects. The MTT assay showed that mitochondrialfunction decreased significantly in cells exposed to the6synthetic pigments, whileMTT values were in a dose-dependent. And there was just a simple addition actionbetween the Acid Red2G and Lissamine Green B, Acid Red2G and Auramine, Acidred2G and Chrysoidine G.