title: Focused ultrasound-assisted acceleration of enzymatic hydrolysis of alkylphenols and 17β-oestradiol glucuronide in fish bile.
abstract: According to the European Water Framework Directive (WFD), alkylphenols, such as octylphenols and nonylphenols, and 17β-oestradiol are considered as priority or emerging pollutants, respectively, mainly due to their possible properties as endocrine-disrupting compounds (EDCs). EDCs are accumulated in liver, fat, kidney and bile in the glucuronide form. In order to determine the concentration of these compounds in bile, an enzymatic hydrolysis step is necessary. This step is usually long (~16 h), and in this sense, ultrasound probes were studied as a possible alternative energy source to accelerate this process. Enzymatic hydrolysis was reduced to 20 min using an ultrasound probe at one cycle and 10% of amplitude. For validation of analytical procedure, nonylphenol glucuronide (4NP-G), 4-tert-octylphenol glucuronide (4tOP-G) and 4-n-octylphenol glucuronide (4nOP-G) were synthesised while 17β-oestradiol glucuronide (E2-G) was commercially available. Bile from thick-lip grey mullets (Chelon labrosus) was spiked with known amounts of 4NP-G, 4tOP-G, 4nOP-G and E2-G and submitted to the optimised procedure. Good recoveries (77-122%), precision in the 5% to 12% range and limits of detection, ranging from the low nanogramme per gramme level for 4tOP, 4nOP and E2 to the low microgramme per gramme level for nonylphenols, were obtained. The optimised method was applied for the determination of alkylphenol in the bile of thick-lip grey mullets fish bile from the Urdaibai estuary (UNESCO reserve of the Biosphere, Bay of Biscay), and high concentrations (2.3-14.2 μg/g), such as those obtained in polluted areas, were measured. E2 was determined in the bile of thick-lip grey mullets, intraperitoneally injected with E2.
authors.affiliation: Department of Analytical Chemistry, University of Basque Country, 644 PK, 48080 Bilbao, Spain. asier.vallejo@ehu.es
authors.forename: Asier
authors.initials: A
authors.lastname: Vallejo
authors.forename: Aresatz
authors.initials: A
authors.lastname: Usobiaga
authors.forename: Maren
authors.initials: M
authors.lastname: Ortiz-Zarragoitia
authors.forename: Miren P
authors.initials: MP
authors.lastname: Cajaraville
authors.forename: Luis A
authors.initials: LA
authors.lastname: Fernández
authors.forename: Olatz
authors.initials: O
authors.lastname: Zuloaga
mesh_terms: D000818:Animals; D001646:Bile / Q000737:chemistry / Q000201:enzymology*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004958:Estradiol / Q000031:analogs & derivatives* / Q000032:analysis / Q000378:metabolism; D005399:Fishes; D006868:Hydrolysis; D013058:Mass Spectrometry / Q000379:methods; D010636:Phenols / Q000032:analysis / Q000378:metabolism*; D014465:Ultrasonics
chemical_list: D010636:Phenols; C025483:estradiol-17 beta-glucuronide; D004958:Estradiol