Validation of a method for the determination of pigments in feed

Identifier:  TFG:127

Handle: https://hdl.handle.net/20.500.11797/TFG127

Authors:  Moreno Gómez, Dulcenombre de Maria

Abstract:
The objective of this work is to develop and validate a quantitative method of analysis for the determination of trans-lutein and trans-canthaxanthin in feed. Both lutein and canthaxanthin are pigments, called carotenoids that belong to the xanthophylls family. Carotenoids have a spectrum in the visible rang of the electromagnetic spectrum that is used for their determination. The method used previously consisted in pigment determinationusing UV-visible spectroscopy. This was an approximated method since all pigments in feed absorb in this range of wavelength and the method is not selective. Then, the determination of one of the carotenoids in a mixture is difficult and we need to apply other techniques like HPLC to have an approximate value of the target analyte. The method used and validated in this work is based on HPLC which allows us to determinate the analytes separately. Due to HPLC has a DAD detector the analytical method will be able to determinate every single pigment that we want to quantify in their maximum of absorbance. As carotenoid standards are expensive, it is applied a method that uses a factor that correlates the response of the cheapest pigment with the rest in the mixture. We quantify the target pigment with the calibration curve of beta-carotene and with the response factor we get the different pigment concentrations.