Annex White book.pdf

Phase Duration (min) Phase A Phase B Phase CEquilibrium 10 90 10 01 0 90 10 02 7 48 52 03 10 48 52 04 14 0 60 405 24 0 60 406 25 90 10 0Table 11. SeparationdevelopmentThe possible artificial dye substances existing in the solution areidentified by comparing the absorption time and the UV-Vis spectrumsbetween 300 and 700 nm with those of the dissolution pattern.The results are described in one decimal mg / kg. Results describedas “present” or “absent” with no limiting detection indication or amountare not acceptable.A4.3.3. Fat-soluble artificial dye substances detection (HPLC)Apart from the ISO/TS 3632 HPLC method applied in Spain, anotherone is also used, allowing the determination of a number of fat-solubleartificial dye substances that belong to the Sudan red family (Sudan I,Sudan II, Sudan III, Sudan IV, 7B Sudan red and Sudan G). They arelargely used for coloring plastics and other synthetic substances. Theyare not indicated for consumption because their “azo-” groups mayeasily transform to carcinogenic amines. In 2003, some of the abovesubstances were detected in a number of food products that containedchili powder. For this reason, the European community is alerted tothe possibility of adding these substances to various spices. The alertexpanded in 2004 on the substances Sudan II, III and IV.The procedure consists in extracting dye substances through acetonitrilefor further chromatographic analysis through HPLC (in reversephase columns and diode array detector). The extraction takes placeafter adding 25 ml of acetonitrile to 500 mg of saffron. The mixture isagitated for 1 minute in a Polytron homogenization apparatus and isfiltered initially through a paper filter and afterwards through polytetraflouroethylene(PTFE) filter with a 0,45 μm pore diameter. The extract isinjected into the HPLC apparatus, which is modified as follows:- Column: C18, length: 250 mm, inner diameter: 4 mm and particlesize: 5 μm.165