High performance liquid chromatographic separation of anabolic oestrogens and ultraviolet detection of 17β‐oestradiol, zeranol, diethylstilboestrol or zearalenone in avian muscle tissue extracts

Abstract

A multi‐residue analysis was developed to discern oestrogens (17β‐oestradiol or zeranol) used as anabolic drugs and diethylstilboestrol (DES) from the mycotoxins zearalenone and zearalenol. A mixture of 17β‐oestradiol, oestrone, DES, zeranol, zearalenol, zearalenone and zearalanone was analysed using a 5 μm silica column with gradient elution from hexane to hexane: methanol: 2‐propanol, 40:45:15 (v/v) and ultraviolet detection at 280 nm. Only six peaks were obtained since zearalenone and zearalanone had identical elution times. Minimum detectabilities were 5–10 ng for the standards. Chicken muscle tissues (2–5 g) were extracted with acetone:water (95:5, v/v). The extracts were spiked with oestradiol and zeranol at 50, 100 or 200 ppb; and DES or zearalenone at 50 ppb; cleaned‐up through dual columns of basic alumina and phosphate exchanged AGMP‐1 resin and analysed by high‐performance liquid chromatography with ultraviolet detection (HPLC‐UV). Minimum detectability was 10 ng for oestradiol, zeranol, DES or zearalenone when tissues were spiked at 50 ppb. Recoveries were ≥86%, ≥83%, 60% and 100% for oestradiol, zeranol, DES and zearalenone, respectively.