Enhanced methods for conditioning, storage, and extraction of liquid and solid samples of manure for determination of steroid hormones by solid-phase extraction and gas chromatography–mass spectrometry

Abstract

Hormones are among the highest-impact endocrine disrupters affecting living organisms in aquatic environments. These molecules have been measured in both wastewater and sewage sludge. Analytical techniques for such matrices are well described in the literature. In contrast, there is little information about the analysis of hormones in animal waste. The objectives of this study were, first, to propose a method for conditioning swine manure samples (addition of formaldehyde, separation of the solid and liquid phases, and duration of storage) in order to determine hormones in the liquid fraction of manure by solid-phase extraction (SPE) coupled with gas chromatography-mass spectrometry (GC-MS). Our results showed that analysis of hormones was affected by matrix changes which occurred during freezing and thawing and after addition of formaldehyde, an additive frequently used to preserve environmental samples. Thus, our results argue for the conditioning of samples without formaldehyde and for separating the solid and liquid fractions of manure before freezing. Second, this study reports on the use of a liquid extraction method coupled with SPE and GC-MS analysis for determination of hormones in the solid fraction of manure. Under the conditions selected, hormone recoveries were between 80 and 100%. Finally, the optimized method was used to quantify hormones in both liquid and solid fractions of swine manure from different breeding units. High levels of estrone and α-estradiol were found in samples whereas β-estradiol was detected in smaller amounts. Estriol and progesterone were mainly found in manure from the gestating sow building whereas testosterone was detected in manure from male breeding buildings.

Androgens and androgenic activity in broiler manure assessed by means of chemical analyses and in vitro bioassays

“…To our knowledge, the present study is the first to report on the presence of DHT in broiler manure. Concentrations of A-dione were previously detected in broiler manure, ranging from 39 ng/g to 96 ng/g [9]. In the present study, levels of A-dione in the manure ranged between 42.49 AE 1.64 ng/g and 503.82 AE 85.2 ng/g.…”

“…However, the chemical analysis revealed thatthe extraction was not asefficient for DHT, A-11β-ol-dione, and Adrfrom manure samples. To our knowledge, this is the first study to report on the presence of DHT in broiler manure.A-dione was previously detected in broiler manure at concentrations ranging from 39 to 96 ng/g [9]. In the present study, levels of A-dione in the manure ranged between 42.49 ± 1.64 and 503.82 ± 85.2ng/g.…”

“…All rights reserved of all farms and in the first sampling of one.Previous studies reported T concentrations of up to 53 ng/g in broiler manure [9].…”

“…To date, analytical chemistry techniques have been the tool of choice in most studies examining the occurrence, fate, and degradation of steroids in manure and soil [8][9][10]. Only a few have applied bioassays based on the use of cell lines able to detect hormonal activities associated with the substances of interest [11][12][13].…”

Centrifuge microextraction coupled with sweeping–MEKC to analyze trace steroid hormones in urine samples

“…Numerous methods have been adopted for the characterization and determination of steroids, such as immunoassay [4,5], gas chromatography-mass spectrometry (GC-MS) [6,7], high performance liquid chromatography-ultra violet detection (HPLC-UV) [8], high performance liquid chromatography-mass spectrometry (HPLC-MS) [9][10][11], capillary electrophoresis-ultra violet detec-tion (CE-UV) [12,13], and capillary electrophoresis-immunoassay [14]. However, rapid and reliable methods are still needed to simplify sample pretreatment and decrease analysis time for highthroughput detection.…”