In vitro biotransformation assays using fish liver cells: Comparing rainbow trout and carp hepatocytes.

Biotransformation assays using primary hepatocytes from rainbow trout, Oncorhynchus mykiss, were validated as a reliable in vitro tool to predict in vivo bioconcentration factors (BCF) of chemicals in fish. Given the pronounced interspecies differences of chemical biotransformation, the present study aimed to compare biotransformation rate values and BCF predictions obtained with hepatocytes from the cold-water species, rainbow trout, to data obtained with hepatocytes of the warm-water species, common carp (Cyprinus carpio). In a first step, we adapted the protocol for the trout hepatocyte assay, including the cryopreservation method, to carp hepatocytes. The successful adaptation serves as proof of principle that the in vitro hepatocyte biotransformation assays can be technically transferred across fish species. In a second step, we compared the in vitro intrinsic clearance rates (CLin vitro, int) of two model xenobiotics, benzo[a]pyrene (BaP) and methoxychlor (MXC), in trout and carp hepatocytes. The in vitro data were used to predict in vivo biotransformation rate constants (kB) and BCFs, which were then compared to measured in vivo kB and BCF values. The CLin vitro, int values of BaP and MXC did not differ significantly between trout and carp hepatocytes, but the predicted BCF values were significantly higher in trout than in carp. In contrast, the measured in vivo BCF values did not differ significantly between the two species. A possible explanation of this discrepancy is that the existing in vitro-in vivo prediction models are parameterized only for trout but not for carp. Therefore, future research needs to develop species-specific extrapolation models

Detection of tetrabromobisphenol A and its mono- and dimethyl derivatives in fish, sediment and suspended particulate matter from European freshwaters and estuaries

An analytical method was developed for the determination of tetrabromobisphenol A (TBBPA), 3,3′,5,5′-tetrabromobisphenol-A-monomethyl ether (MM-TBBPA) and 3,3′,5,5′-tetrabromobisphenol-A-dimethyl ether (DM-TBBPA), and its valid application on fish muscle matrix (bream and sole), suspended particulate matter (SPM) and surface sediment layer samples, using only 0.5 g sample material, is demonstrated. Here, for the first time, DM-TBBPA could be determined by an LC-MS/MS-based method applying atmospheric pressure photoionization (APPI), using the same sample extracts for all three analytes. Samplings covered freshwater fish (bream; annually, period 2007–2013) and SPM or sediment (every second year in the period 2008–2014) at selected European sites (rivers: Tees/UK, Mersey/UK, Western Scheldt/NL, Götaälv/SE, Rhône/FR; Lake Belau/DE). TBBPA could be quantified in 13 of 36 bream samples (range about 0.5–1.2 μg kg−1 ww) and 7 of 7 sole muscle samples (range about 0.5–0.7 μg kg−1 ww). Further, it could be quantified in 11 of the 14 SPM samples (range about 0.5–9.4 μg kg−1 dw) and in both of the surface sediment layer samples (2.3–2.6 μg kg−1 dw). MM-TBBPA could be quantified in 12 of 36 bream and 4 of 7 sole muscle samples (range about 0.8–1.8 μg kg−1 ww). Further, it could be quantified in 10 of the 14 river SPM samples (range about 2.3–4.5 μg kg−1 dw) and in both lake surface sediment layer samples (5.2–5.5 μg kg−1 dw). DM-TBBPA was rarely detectable and could not be quantified above the limit of quantification in any sample

DataSheet1_In vitro biotransformation assays using fish liver cells: Comparing rainbow trout and carp hepatocytes.pdf

Biotransformation assays using primary hepatocytes from rainbow trout, Oncorhynchus mykiss, were validated as a reliable in vitro tool to predict in vivo bioconcentration factors (BCF) of chemicals in fish. Given the pronounced interspecies differences of chemical biotransformation, the present study aimed to compare biotransformation rate values and BCF predictions obtained with hepatocytes from the cold-water species, rainbow trout, to data obtained with hepatocytes of the warm-water species, common carp (Cyprinus carpio). In a first step, we adapted the protocol for the trout hepatocyte assay, including the cryopreservation method, to carp hepatocytes. The successful adaptation serves as proof of principle that the in vitro hepatocyte biotransformation assays can be technically transferred across fish species. In a second step, we compared the in vitro intrinsic clearance rates (CLin vitro, int) of two model xenobiotics, benzo[a]pyrene (BaP) and methoxychlor (MXC), in trout and carp hepatocytes. The in vitro data were used to predict in vivo biotransformation rate constants (kB) and BCFs, which were then compared to measured in vivo kB and BCF values. The CLin vitro, int values of BaP and MXC did not differ significantly between trout and carp hepatocytes, but the predicted BCF values were significantly higher in trout than in carp. In contrast, the measured in vivo BCF values did not differ significantly between the two species. A possible explanation of this discrepancy is that the existing in vitro-in vivo prediction models are parameterized only for trout but not for carp. Therefore, future research needs to develop species-specific extrapolation models.</p