Leif Kronberg

Determination of the strong mutagen 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone in chlorinated drinking and humic waters☆

The strong Ames test mutagen 3-chloro-4-(dichloromethyl)-5-hydroxy-2-(5H)-furanone (MX) was found to occur in chlorinated drinking and humic waters in sub μg L−1 concentrations. Quantitation by GC/MS using Selective Ion Monitoring in parallel with Ames tests of waters indicated that this furanone accounted for 15 – 30% of the Ames test mutagenicity of both a drinking water and a humic water.

Ames mutagenicity and concentration of the strong mutagen 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone and of its geometric isomer E-2-chloro-3-(dichloromethyl)-4-oxo-butenoic acid in chlorine-treated tap waters

The Ames mutagenicity and the concentration of the strong Ames mutagen 3-chloro-4-(dichloro-methyl)-5-hydroxy-2(5H)-furanone (MX) and its geometric isomer E-2-chloro-3-(dichloromethyl)-4-oxobutenoic acid (E-MX), derived from chlorination of humus, were determined in XAD extracts of tap water collected from 26 localities in Finland. The 23 tap waters treated with disinfectants gave a positive response in strain TA100. MX and E-MX were detected in all extracts exhibiting mutagenicity with the exception of 3 extracts of marginal activity. MX accounted for 15-57% (average 33%) of the observed mutagenicity. The concentration of E-MX was slightly lower than the corresponding concentration of MX. Linear correlations were observed between mutagenicity and concentration of MX and E-MX, with correlation coefficients of 0.894 for MX and 0.910 for E-MX.

Identification and quantification of the Ames mutagenic compound 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone and of its geometric isomer (E)-2-chloro-3-(dichloromethyl)-4-oxobutenoic acid in chlorine-treated humic water and drinking water extracts.

Mutagenic compounds in XAD extracts of chlorinated humic water were separated in two stages of fractionation by reversed-phase high-performance liquid chromatography. Analyses of mutagenic fractions by gas chromatography/mass spectroscopy resulted in the identification of the strong Ames mutagen 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone (MX) and its geometric isomer (E)-2-chloro-3-(dichloromethyl)-4-oxobutenoic acid (E-MX). Both compounds were also detected in extracts of chlorinated drinking waters. MX accounted for 50-100% of the activity of extracts of chlorinated humic water and for 20-50% of the activity of extracts of drinking water. E-MX exhibited at most one-tenth of the MX mutagenicity and accounted for 2% or less of the activity of the extracts of drinking water. However, because of the poor reproducibility of E-MX mutagenicity tests and the ability of the compound to isomerize the MX, there is uncertainty about the significance of E-MX in chlorinated waters.

Inkjet printing of drug substances and use of porous substrates‐towards individualized dosing

Medicines are most often oral solid dosage forms made into tablets or capsules, and there is little room for individualized doses. The drug substance and additives are processed through multiple production phases, including complex powder handling steps. In drug manufacturing, the control of the solid-state properties of active pharmaceutical ingredient (API) is essential and it offers opportunities for enhancement of drug delivery systems. In this context, inkjet printing technologies have emerged over the last decades in pharmaceutical and biological applications and offer solutions for controlling material and product characteristics with high precision. Here we report the concept of conventional inkjet printing technology to produce printable pharmaceutical dosage forms on porous substrates. Data are shown to demonstrate inkjet printing of APIs into paper substrates, and how the model drug substances (paracetamol, theophylline, and caffeine) are penetrating the porous substrates used. The method enables controlling not only the deposition but also the crystallization of the drug substances. We anticipate that the inkjet printing approach has immense potential in making sophisticated drug delivery systems by use of porous substrates in the future. For example, it may offer new perspectives for solving problems around poorly soluble drugs and dosing low-dose medicines accurately. Furthermore, with the advent of genetic mapping of humans, controlled inkjet dosing can bring solutions to fabricate on-demand individualized medicines for patients.

The anti-inflammatory drugs diclofenac, naproxen and ibuprofen are found in the bile of wild fish caught downstream of a wastewater treatment plant.

Pharmaceutical residues are ubiquitous in rivers, lakes, and at coastal waters affected by discharges from municipal wastewater treatment plants. In this study, the presence of 17 different pharmaceuticals and six different phase I metabolites was determined in the bile of two wild fish species, bream (Abramis brama) and roach (Rutilus rutilus). The fish were caught from a lake that receives treated municipal wastewater via a small river. Prior to analyses, the bile content was enzymatically hydrolyzed to convert the glucuronide metabolites into the original pharmaceuticals or phase I metabolites. The solid phase extracts of hydrolyzates were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in the multiple reaction monitoring mode. The anti-inflammatory drug naproxen could be detected in all the six bream and roach bile samples. Diclofenac was found in five of the bream and roach samples, while ibuprofen was detected in three bream and two roach samples. The observed bile concentrations of diclofenac, naproxen, and ibuprofen in bream ranged from 6 to 95 ng mL(-1), 6 to 32 ng mL(-1), and 16 to 34 ng mL(-1), respectively. The corresponding values in roach samples ranged from 44 to 148 ng mL(-1), 11 to 103 ng mL(-1) and 15 to 26 ng mL(-1), respectively. None of the other studied compounds could be detected. The study shows that pharmaceuticals originating from wastewater treatment plant effluents can be traced to the bile of wild bream and roach living in a lake where diclofenac, naproxen, and ibuprofen are present as pollutants.

Seasonal variations in the occurrence and fate of basic and neutral pharmaceuticals in a Swedish river–lake system

The seasonal variations in the occurrence of carbamazepine, atenolol, metoprolol, sotalol, and acebutolol were studied at seven sites along River Fyris from December 2007 to December 2008. Samples were collected from the effluent of a waste water treatment plant (WWTP), at one upstream site, and five downstream sites of the WWTP. During one occasion in May 2008, water samples were collected at different locations and depths in the recipient lake. All analytes except of acebutolol were present in both the river and the lake at quantifiable amounts at all sampling occasions. Carbamazepine was found in similar concentrations (about 90 ng L(-1)) at all sampling sites and all studied depths (0.5-40 m) in the lake, indicating high environmental persistence of this compound. A clear seasonal pattern was observed for the natural attenuation of the beta-blockers in the river, with the highest attenuation occurring in summer and the lowest in winter. The loss of beta-blockers on a distance of 1320 m reached up to 75% during summer time but was insignificant during winter. The seasonal variations in the loss followed the seasonal variations in water temperature and chlorophyll a mass flow suggesting that biotransformation and adsorption are the main processes responsible for the loss of the studied pharmaceuticals in River Fyris downstream the WWTP.

Removal of Pharmaceuticals in Drinking Water Treatment: Effect of Chemical Coagulation

The removal of selected pharmaceuticals (diclofenac, ibuprofen, bezafibrate, carbamazepine and sulfamethoxazole) by chemical coagulation was studied. Jar test experiments were done in MilliQ water, in lake water and in commercial humic acid solutions using aluminium (pH 6) and ferric sulphate (pH 4.5). The concentrations of the pharmaceuticals in the studied water samples were determined by HPLC analysis and UV detection. In MilliQ water coagulation, the pharmaceuticals were poorly removed (< 10%) with the exception of diclofenac, which was removed up to 66% with ferric sulphate. This compound was also the only pharmaceutical removed (30%) during the lake water coagulation with ferric sulphate. In the presence of dissolved humic matter, diclofenac as well as ibuprofen and bezafibrate could be removed by ferric sulphate coagulation. The removal of diclofenac reached a maximum of 77%, while 50% of ibuprofen and 36% of bezafibrate were removed. Hence, a high amount of high-molecular-weight dissolved organic matter enhanced the removal of ionisable pharmaceuticals. The non-ionisable compounds, carbamazepine and sulfamethoxazole, were not affected by the coagulation processes studied. Although conditions such as high humic material content, low coagulation pH and ferric coagulant increase the removal of certain ionic pharmaceuticals, it can be concluded that by coagulation it is not possible to entirely remove pharmaceuticals from water.

Uptake from water, biotransformation, and biliary excretion of pharmaceuticals by rainbow trout

An urgent need exists to assess the exposure of fish to pharmaceuticals. The aim of the present study was to assess the uptake and metabolism of waterborne pharmaceuticals in rainbow trout (Oncorhynchus mykiss). A further objective was to determine the possibility of monitoring exposure to low levels of pharmaceuticals by bile assays. Rainbow trout were exposed for 10 d under flow-through conditions to mixtures of five pharmaceuticals (diclofenac, naproxen, ibuprofen, bisoprolol, and carbamazepine) at high and low concentrations. The low concentration was used to mimic the conditions prevailing in the vicinity of the discharge points of wastewater treatment plants. The uptake and the bioconcentration were determined by blood plasma and bile analyses. The average bioconcentration factor in plasma ranged from below 0.1 for bisoprolol to 4.9 for diclofenac, the values being approximately similar at low and high ambient concentrations. The biotransformation of diclofenac, naproxen, and ibuprofen was considered efficient, because several metabolites could be detected in concentrations clearly exceeding those of the unmetabolized compounds. The glucuronides were the dominant metabolites for all three pharmaceuticals. The total bioconcentration in the bile was two to four orders of magnitude higher than in the plasma. The results of this work show that the exposure of fish to pharmaceuticals in environmentally relevant concentrations may be monitored by blood plasma and bile analyses, the latter allowing detection at markedly lower ambient concentration.

Metabolites of the Aquatic Pollutant Diclofenac in Fish Bile

The uptake and metabolism of anti-inflammatory drug diclofenac (DCF) was studied by exposing rainbow trout (Oncorhynchus mykiss) to DCF intraperitoneally, and via water at concentration of 1.7 μg L(-1). The bile was collected and the formed metabolites were identified. The identification was based on the exact mass determinations by a time-of-flight mass analyzer and on the studies of fragments and fragmentation patterns of precursor ions by an ion trap mass analyzer. The main metabolites found were acyl glucuronides of hydroxylated DCFs. In addition, one ether glucuronide of hydroxylated DCF was found. Also, unmetabolized DCF was detected in the bile. The total bioconcentration factors (BCF(total-bile) for DCF and its metabolites) in rainbow trout bile, varied between individuals and was roughly estimated to range from 320 to 950. These findings suggest that fish living downstream the wastewater treatment plants (WWTPs) and which are chronically exposed to the drug may accumulate the drug and its metabolites in the bile.

Oligolignans in Norway spruce and Scots pine knots and Norway spruce stemwood

Abstract Oligolignans present in substantial amounts in Norway spruce and Scots pine knots were characterised. The hydrophilic knotwood substances were extracted and different chromatographic methods were applied to obtain fractions for analysis by GC, GC-MS, HR-EI-MS, LC-ESI-MS, NMR, and FTIR. β-O-4-Linked guaiacylglyceryl ethers of hydroxymatairesinol (Ia, Ib, Ic), secoisolariciresinol (II), lariciresinol (III), isolariciresinol (IV), lignan A (V), liovil (VI), conidendrin (VII), and pinoresinol (VIII) were identified in the spruce knotwood extract and β-O-4-linked guaiacylglyceryl ethers of nortrachelogenin (IX) and secoisolariciresinol (II) in the pine knotwood extract. The structures of allo-hydroxymatairesinol-4′-guaiacylglyceryl ether (Ia), hydroxymatairesinol-4-guaiacylglyceryl ether (Ib), hydroxymatairesinol-4′-guaiacylglyceryl ether (Ic), secoisolariciresisinol-4-guaiacylglyceryl ether (II), and lariciresinol-4′-guaiacylglyceryl ether (III) were determined via GC-MS, HR-MS, and 1H and 13C NMR analyses. The structures of (Ia), (Ic), (IV), (V), (VI), (VII), and (IX) have not been reported earlier. Spruce stemwood, both heartwood and sapwood, also contained oligolignans in small amounts. In addition, some dilignans with four phenylpropanoid units were tentatively identified in the hydrophilic knotwood substances.