Development and validation of a high performance liquid chromatography/diode array detection method for estrogen determination: Application to residual analysis in meat products

In this work, an HPLC-DAD method was developed for the residual analysis of some estrogens such as estrone (E1), 17-β estradiol (E2), estriol (E3), natural estrogens, and 17-α ethinylestradiol (E4), an exoestrogen, in meat samples of different categories (chicken, n = 155, beef, n = 124, sheep, n = 122, and camels, n = 40), collected from the Saudi Market. Although banned, the use of E4 as a growth promoter in the black market is still encountered. Symmetry C18 column (3.5 µm, 4.6 mm × 150 mm) was used with a mobile phase consisting of 50% aqueous acetonitrile. Protein precipitation with acetonitrile was used for the sample preparation. The method was fully validated, as per the ICH guidelines, in the concentration ranges of 0.35–125 µg/g (E1, E2), 0.188–125 µg/g (E3), and 0.188–450 µg/g (E4). The method allowed the trace analysis of estrogens with LOD values of 0.094 (E3, E4) and 0.126 µg/g (E1, E2), and LOQ values of 0.188 (E3, E4) and 0.350 µg/g (E1, E2). The analyzed samples contained different levels of estrogens. Within the same category, processed products contained the highest levels of E4, while the internal organs contained the least estrogen content. Finally, the estimated daily intake, µg/kg bw/day, of estrogens through the consumption of meat-based food products was calculated