Abstract
In term of pharmaceutical and their intermediate compounds analysis, UPLC/MS method is a valuable equipment to achieve better confirmation on their biodegradation by fungi. The T47D-KBluc reporter gene assay is an appropriate tool to investigate to removal of estrogenic and antiestrogenic activities of pharmaceuticals and their metabolites from a synthetic wastewater. A consortium of isolated South African indigenous fungi Aspergillus niger, Mucor circinelloides, Trichoderma longibrachiatum, Trametes polyzona and Rhizopus microspores was found to perform a removal of pharmaceuticals and their metabolites and to reduce their estrogenic activity below the limit of detection in a sequencing batch reactor. Here are presented data regarding the phenolic compounds list and the method validation for UPLC/MS analysis used for selected pharmaceutical compounds namely carbamazepine, diclofenac, ibuprofen and their metabolites, as well as the T47D-KBluc bioassay using as positive control, the agonist E2 for estrogenic activity and the antagonist ICI 182,780 for antiestrogenic activity. For better understanding of the data presented in this paper, please see the research paper “Removal of pharmaceutical’ estrogenic activity of sequencing batch reactor effluents assessed in the T47DK-Bluc reporter gene assay” [1].
Highlights
In term of pharmaceutical and their intermediate compounds analysis, Ultra-Performance Liquid Chromatography tandem Mass Spectrometry (UPLC/MS) method is a valuable equipment to achieve better confirmation on their biodegradation by fungi
Ultra-Performance Liquid Chromatography Water Acquity UPLC® system hyphenated to a quadrupole-time-of-flight (QToF) instrument operating with MassLynxTM software (Waters Inc., Milford, Massachusetts, USA) was used for data acquisition and processing
Estrogenic and antiestrogenic activity was calculated from relative light unit using LUMIstar OPTIMA luminometer (BMG Labtech, Germany) Raw and Analysed Solid phase extraction (SPE) method using SupelTM Select Supelco HLB 500 mg, 12 mL plastic cartridges (Sigma Aldrich, South Africa) was performed for UPLC/MS samples loaded at pH 2.5, eluted with methanol (HPLC grade, Sigma Aldrich, South Africa)
Summary
The T47D-KBluc reporter gene assay method was developed by the United States Environmental Protection Agency (USEPA) to screen chemical compounds and environmental samples for estrogenic and anti-estrogenic activities. T47D human breast cancer cells, which naturally express both human estrogen receptor (ER)-a and -b, were transferred with an estrogen-responsive element luciferase reporter gene construct [5]. The 17-b estradiol (E2) agonist positive control is a natural steroid sex hormone produced by the ovaries named according to UIPAC as (8R,9S, 13S, 14S, 17S)-13-methyl6,7,8,9,11,12,14,15,16,17-decahydrocyclopenta[a]phenanthrene-3,17-diol [1], with molecular mass of 272.388 g/mol. The antagonist ICI 182,780 is a synthetic steroidal antiestrogen derived from E2 named by IUPAC as (7R, 8R, 9S, 13S, 14S, 17S)-13-methyl-7-[9-(4,4,5,5,5-pentafluoropentylsulfinyl)nonyl]6,7,8,9,11,12,14,15, 16,17-decahydrocyclopenta[a]phenanthrene-3,17-diol [1], with a molecular mass of 606.777 g/mol
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