Courtesy of the Royal Society of Chemistry
Figure 45 Chromatogram of Bixin and Nor bixin Contained in Plasma After Ingestion of Annato Food Colors
The extraction process was repeated three times and the three extracts combined. The bulked extracts were evaporated to dryness and the residue mixed with one ml of methanol sonicated and the supernatant liquid used for analysis.
The column used was Spherisorb S60DS1 (reversed phase) and a mobile phase consisting of a mixture of 70%v/v, acetonitrile 30%v/v aqueous 2% acetic acid at a flow rate of 1.5 m;l/min. The sample volume used was 10 ml and the solution of the standard (0.8mg/ml) was made by dissolving 4 mg of Sudan 1 in 100 ml of methanol and diluting one ml of the solution to 50 ml with methanol. A typical example of the separation is shown in figure 45.The quantity of bixin and norbixin isomers was obtained from a chromatogram of the standard Sudan 1, which is shown in figure 46. The same technique was used to examine the metabolism of bixin compounds over different ingestion periods. The chromatograms obtained from the analysis of serum 2 hrs and 4 hrs after ingestion are shown in figure 47.
Courtesy of the Royal Society of Chemistry
Figure 46 Chromatogram of Annatto Food Color Used for Ingestion
