Stability, preservation, and quantification of hormones and estrogenic and androgenic activities in surface water...

Degradation of hormones that may occur during storage of surface water samples can lead to underestimations in estrogenic and androgenic activities and inaccuracies in hormone concentrations. The current study investigated the use of sodium azide, hydrochloric acid (HCl), and sulfuric acid (H₂SO₄) to inhibit the degradation of hormones and estrogenic and androgenic activities in samples of surface water runoff from cattle manure-amended fields during storage at 4°C. Hormones and hormone metabolites were extracted using solid-phase extraction and analyzed using high-performance liquid chromatography (HPLC) with tandem MS. Estrogenic and androgenic activities were assessed by E-screen and A-screen, respectively. Results of the current study indicate significant degradation of estrogenic, androgenic, and progestogenic hormones and activities, which is likely attributable to microbial activity, within hours of sample collection. The inclusion of internal standards provides a means to account for hormone losses caused by extraction inefficiency and to some extent degradation. However, internal standards are unable to adequately account for significant losses and are not available for E-screen and A-screen. Sodium azide did not adequately inhibit androgen degradation at the concentration used (1 g/L). Acid preservation (HCl or H₂SO₄, pH 2) stabilized the estrogenic and androgenic activities, and coupling acid preservation with the use of internal standards resulted in reliable and accurate recovery of a suite of androgens, estrogens, and progestogens for up to 14 d of storage at 4°C.