High resolution mass spectrometric identification and quantification of glucocorticoid compounds in various wastewaters in The Netherlands

In the past two decades much research effort has focused on the occurrence, effects and risks of
estrogenic compounds. However, increasing emissions of new emerging compounds may also affect the
action of hormonal pathways other than the estrogenic hormonal axis. Recently, a suite of novel CALUX®
bioassays has become available that enables to look further than estrogenic effects only. By employing
these bioassays, we recently showed high glucocorticogenic activity in wastewaters collected at various
sites in The Netherlands. However, since bioassays provide an integrated biological response, the
identity of the responsible biological compounds remained unknown. Therefore, our current objective
was to elucidate the chemical composition of the wastewater extracts used in our previous study by
means of LC-high resolution Orbitrap® MS/MS and to determine if the compounds quantified could
account for the observed glucocorticoid responsive (GR) CALUX bioassay response. The mass
spectrometric analysis revealed the presence of various glucocorticoids in the range of 13-1900 ng/L. In
extracts of hospital wastewater – collected prior to sewage treatment – several glucocorticoids were
identified (cortisol: 275-301 ng/L, cortisone: 381-472 ng/L, prednisone: 117-545 ng/L, prednisolone: 315-
1918 ng/L and triamcinolone acetonide: 14-41 ng/L) which are used to treat a great number of human
pathologies. A potency balance calculation based on the instrumental analyses and relative potencies
(REPs) of the individual glucocorticoids, supports the conclusion that triamcinolone acetonide (REP=1.3),
dexamethasone (REP=1) and prednisolone (REP=0.2) are the main contributors to the glucocorticogenic
activity in the investigated wastewater extracts. The action of these compounds is concentration additive
and the overall glucocorticogenic activity can be explained to a fairly large extent by their contribution.