Point-of-care therapeutic drug monitoring of tumour necrosis factor-α inhibitors using a single step immunoassay.

Therapeutic drug monitoring (TDM) of tumor necrosis factor-α (TNFα)-inhibitors adalimumab and infliximab is important to establish optimal drug dose and maximize treatment efficacy. Currently, TDM is primarily performed with ELISA techniques in clinical laboratories, resulting in a long sample-to-result workflow. Point-of-care (POC) detection of these therapeutic antibodies could significantly decrease turnaround times and allow for user-friendly home-testing. Here, we adapted the recently developed bioluminescent dRAPPID (dimeric Ratiometric Plug-and-Play Immunodiagnostics) sensor platform to allow POC TDM of infliximab and adalimumab. We applied the two best performing dRAPPID sensors, with limit-of-detections of 1 pM and 17 pM, to measure the infliximab and adalimumab levels in 49 and 40 patient serum samples, respectively. The analytical performance of dRAPPID was benchmarked with commercial ELISAs and yielded Pearson's correlation coefficients of 0.93 and 0.94 for infliximab and adalimumab, respectively. Furthermore, a dedicated bioluminescence reader was fabricated and used as a readout device for the TDM dRAPPID sensors. Subsequently, infliximab and adalimumab patient serum samples were measured with the TDM dRAPPID sensors and bioluminescence reader, yielding Pearson's correlation coefficients of 0.97 and 0.86 for infliximab and adalimumab, respectively, and small proportional differences with ELISA (slope was 0.97 ± 0.09 and 0.96 ± 0.20, respectively). The adalimumab and infliximab dRAPPID sensors, in combination with the dedicated bioluminescence reader, allow for ease-of-use TDM with a fast turnaround time and show potential for POC TDM outside of clinical laboratories.

Determination of chloroacetanilides, triazines and phenylureas and some of their metabolites in soils by pressurised liquid extraction, GC-MS/MS, LC-MS and LC-MS/MS.

Pressurised liquid extraction (PLE) technique was used for the simultaneous extraction of phenylureas, triazines and chloroacetanilides and some of their metabolites from soils. Extractions were performed by mixing 15 g of dried soil with 30 mL of acetone under 100 atm at 50 degrees C, during 3 min and with three PLE cycles. Prior to the analysis of naturally contaminated soils, each of the five representative soil matrices used as blanks (of different depths) was spiked in triplicate with standards of each parent and degradation compound at about 10, 30 and 120 microg/kg. For each experiment, isoproturon-D6 and atrazine-D5 were used as surrogates. Analysis of phenylureas and metabolites of triazines and phenylureas was carried out by reversed phase liquid chromatography/mass spectrometry (LC-MS) and LC-MS/MS in the positive mode. Gas chromatography (GC)/ion trap mass spectrometry was used in the MS/MS mode for the parent triazines and chloroacetanilides. The average extraction recoveries were above 85%, except for didesmethyl-isoproturon, and quantification limits were between 0.5 and 5 microg/kg. The optimised multi-residue method was applied to soils and solids below the root zone, sampled from agricultural plots of a small French hydrogeological basin.

Stability of organophosphorus insecticides on graphitized carbon black extraction cartridges used for large volumes of surface water.

The stability of nine organophosphorus insecticides (azinphos-ethyl, azinphos-methyl, diazinon, EPN, ethion, fonofos, malathion, phosmet and parathion-methyl) was evaluated under a variety of storage conditions. Large volumes of surface water (4 l) were extracted using large-particle-size graphitized carbon black cartridges (Carbopack B 60-80 mesh). The effects of temperature, matrix type and drying of cartridges on the recovery of these contaminants, after different storage periods, were studied and compared to the conservation of surface water in bottles. After a 2-month period, all the chemicals stored on cartridges and kept at -20 degrees C remained stable, with recoveries ranging from 70 to 134%. By contrast, phosmet and EPN could no longer be recovered from the bottled surface water. Cartridges kept at -20 degrees C fared better than did those stored at 4 degrees C and 20 degrees C. The type of matrix water selected appears to have kept the target pesticides stored on cartridges from degrading, compared to the Milli-Q water, in which malathion and phosmet were unstable. The effect of the cartridges being either wet or dry made no difference in terms of improving the recovery of chemicals. After immediate surface water extraction, the most practical storage condition for the target insecticides was found to be storage on cartridges in the dark at -20 degrees C, with no drying or solvent washing of the Carbopack B material.

Determination of organonitrogen pesticides in large volumes of surface water by liquid-liquid and solid-phase extraction using gas chromatography with nitrogen-phosphorus detection and liquid chromatography with atmospheric pressure chemical ionization mass spectrometry.

During a recent study to determine the fluxes and fates of contaminants in the St. Lawrence River, the majority of organonitrogen pesticides analysed in samples of surface water were found in the dissolved phase. This paper compares two extraction techniques and two analytical techniques for 10 chemicals (metolachlor, seven triazines and two degradation products of atrazine-cyanazine-propazine and simazine) in the dissolved phase in large volumes of surface water, using a fibre glass filter with 0.7 micron porosity. Samples of filtered surface water (1-20 l) were extracted by means of a liquid-liquid technique using the Goulden large-sample extractor, and by means of a solid-phase extraction technique, using cartridges filled with 500 mg of a large particle-size graphitized carbon black as adsorbent: Carbopack B (500-666 microns). The pesticides were analysed by gas chromatography on two DB-5 and DB-210 capillary columns with nitrogen-phosphorus detection (GC-NPD) and by liquid chromatography coupled with mass spectrometry equipped with an atmospheric pressure chemical ionization interface (LC-APCI-MS). The recoveries were high (67-100%) for the majority of the target pesticides in a volume of 17.85 l of Milli-Q water, compared to recoveries in the same volume of filtered surface water (51-102%). The detection limits ranged from 0.4 to 4 ng/l and from 0.6 to 3 ng/l for GC-NPD and LC-ACPI-MS techniques, respectively.

Multiresidue methods using solid-phase extraction techniques for monitoring priority pesticides, including triazines and degradation products, in ground and surface waters.

The review describes the use of solid-phase extraction (SPE) techniques for monitoring priority pesticides in ground and surface waters. The focus is on triazine herbicides and their degradation products. Data concerning the fate, occurrence, properties and extraction of triazines and their degradation products using different SPE techniques are tabulated and discussed.

Speciation of butyl- and phenyltin compounds in sediments using pressurized liquid extraction and liquid chromatography-inductively coupled plasma mass spectrometry.

A liquid chromatographic method with inductively coupled plasma mass spectrometry is proposed for the speciation of butyl- (monobutyltin, dibutyltin, tributyltin) and phenyl- (monophenyltin, diphenyltin, triphenyltin) tin compounds in sediments. After evaluation of different additives in the mobile phase, the use of 0.075% (w/v) of tropolone and 0.1% (v/v) of triethylamine in a mobile phase of methanol-acetic acid-water (72.5:6:21.5) allowed the best chromatographic separation of the six compounds. Pressurized liquid extraction (PLE) with a methanolic mixture of 0.5 M acetic acid and 0.2% (w/v) of tropolone was suitable for the quantitative extraction of butyl- and phenyltin compounds with recovery values ranging from 72 to 102%. This analytical approach was compared to conventional solvent extraction methods making use of acids and/or organic solvent of medium polarity. The main advantages of PLE over conventional solvent extraction are: (i) the possibility to extract quantitatively DPhT and MPhT from sediments, which could not be done by a solvent extraction approach; (ii) to preserve the structural integrity of the organotin compounds; (iii) to reduce the extraction time from several hours in case of solvent extraction techniques to just 30 min. For spiked sediments, limits of detection ranged from 0.7 to 2 ng/g of tin according to the compound. The relative standard deviations were found to be between 8 and 15%. The developed analytical procedure was validated using a reference material and was applied to various environmental samples.

Application of liquid chromatography with mass spectrometry combined with photodiode array detection and tandem mass spectrometry for monitoring pesticides in surface waters.

Liquid chromatography with photodiode array detection (LC-DAD) and liquid chromatography with mass spectrometry (LC-MS) are two techniques that have been widely used in monitoring pesticides and their degradation products in the environment. However, the application of liquid chromatography with tandem mass spectrometry (LC-MS-MS) for such purposes, once considered too costly, is now gaining considerable ground. In this study, we compare these methods for the multi-residue analysis of pesticides in surface waters collected from the central and southeastern regions of France, and from the St. Lawrence River in Canada. Forty-eight pesticides belonging to eight different classes (triazine, amide, phenylurea, triazole, triazinone, benzimidazole, morpholine, phenoxyalkanoic), along with some of their degradation products, were monitored on a regular basis in the surface waters. For LC-MS, we used the electrospray ionization (ESI) interface in the negative ionization mode on acidic pesticides (phenoxyalkanoic, sulfonylurea), and the atmospheric pressure chemical ionization (APCI) interface in the positive ionization mode on the remaining chemicals. Different extraction techniques were employed, including liquid-liquid extraction with dichloromethane, and solid-phase extraction using C18-bonded silica and graphitized carbon black cartridges. Eleven of the target chemicals (desethylatrazine, desisopropylatrazine, atrazine, simazine, terbuthylazine, metolachlor, carbendazime, bentazone, penconazole, diuron and isoproturon) were detected by LC-MS at concentrations ranging from 20 to 900 ng/l in the surface waters from France, and six pesticides (atrazine, desethylatrazine, desisopropylatrazine, cyanazine, simazine and metolachlor) were detected by LC-MS and LC-MS-MS at concentrations ranging from 3 to 52 ng/l in the samples drawn from the St. Lawrence River. There was good correlation between the LC-DAD and LC-MS techniques for 60 samples. The slope of the curves expressing the relationship between the results obtained with LC-DAD versus those obtained by LC-MS was near 1, with a correlation coefficient (r) of over 0.93. The identification potential of the LC-MS technique, however, was greater than that of the LC-DAD; its mass spectra, mainly reflecting the pseudomolecular ion resulting from a protonation or a deprotonation of the molecule, was rich in information. The LC-MS-MS technique with ion trap detectors, tested against the LC-MS on 10 surface water samples, gave results that correlated well with the LC-MS results, albeit generating mass spectra that yielded far more information about the structure of unknown substances. The sensitivity of the LC-MS-MS was equivalent to the selected ion monitoring (SIM) acquisition mode in LC-MS. The detection limits of the target pesticides ranged from 20 to 100 ng/l for the LC-MS technique (under full scan acquisition), and from 2 to 6 ng/l for LC-MS-MS. These limits were improved by a factor of almost 10 by increasing the sample volume to 10 l.

Determination of oxanilic and sulfonic acid metabolites of acetochlor in soils by liquid chromatography-electrospray ionisation mass spectrometry.

An analytical method is presented that describes the extraction and quantification of oxanilic and sulfonic acid metabolites of the herbicide acetochlor in soil samples. Experiments were performed on 50 g of soil using a solvent extraction technique with an acetonitrile-water (60:40) mixture in an acidic medium. Analysis was carried out by reversed-phase liquid chromatography and detection by electrospray ionisation mass spectrometry in single ion monitoring and negative modes. Four different soil matrices were spiked in triplicate with standards of each degradation compound at three concentration levels between 2 and 80 microg/kg. The average recoveries range from 90 to 120% for both the metabolites, with relative standard deviations lower than 15%. The limits of quantification are about 1 and 2 microg/kg for the ethanesulfonic acid and the oxanilic acid metabolites, respectively. The method has been applied to soils and solids recovered from the deeper unsaturated zone of a small French catchment closely monitored as part of the European project "Pesticides in European Groundwaters: detailed study of Aquifers and Simulation of possible Evolution scenarios (PEGASE)".

Stability of isoproturon, bentazone, terbuthylazine and alachlor in natural groundwater, surface water and soil water samples stored under laboratory conditions.

The stability of isoproturon, bentazone, terbuthylazine and alachlor was investigated in groundwater (GrW), surface water (SuW) and soil water from the unsaturated zone (SoW). Samples fortified with a low spiking level (LSL) of about 0.3-0.5 microgram/L and a high spiking level (HSL) of about 0.9-1.3 micrograms/L were stored for 1, 2, 14 (GrW) and 30 days (SuW and SoW) at 4 degrees C in amber glass bottles without biological inhibition. The initial pesticide concentration played a significant role, the lowest concentrations being the least stable for all pesticides. Nevertheless, after 14 days of storage, no concentration had decreased significantly compared to day 0 values, except for bentazone LSL in the GrW and SuW. Significant losses of alachlor were observed only after 30 days. Terbuthylazine and isoproturon were stable for 30 days, except for a slight loss of terbuthylazine HSL in the SoW. The very poor recovery of bentazone from the SoW gave poor results for interpretation. Overall, the stability of the molecules was highest in the GrW and lowest in the SoW. For SoW, the variability of triplicate determinations at a given storage time was, in some cases, as great as the changes in mean concentrations observed over the total 30 day storage period.

Occurrence of alkylphenol polyethoxylates in the St. Lawrence River and their bioconcentration by mussels (Elliptio complanata).

A study was conducted in 1999 to determine the occurrence of alkylphenol polyethoxylates in the St. Lawrence River and their bioconcentration by mussels (Elliptio complanata). Concentrations of selected contaminants were measured in surface water, municipal effluent, sediments and mussels. Analyses were performed on 4-tert-octylphenol (4-t-OP), 4-n-nonylphenol (4-n-NP), nonylphenol polyethoxylates (NP(1-16)EO), nonylphenol-mono and di-ethoxycarboxylic acids (NP(1)EC and NP(2)EC), and octylphenol-mono and di-ethoxycarboxylic acids (OP(1)EC and OP(2)EC). Mussels (Elliptio complanata) taken from a reference lake were placed in cages and submerged for 62 days at two sites in the St. Lawrence River, 1.5 km upstream and 5 km downstream of the outfall of a municipal wastewater treatment plant. The results showed that many of the target chemicals were present in all matrices studied: in water, at ppt and ppb levels, and reaching ppm levels in sediments and mussels. Concentrations of these contaminants were higher in matrices sampled at the downstream site than in those drawn at the site upstream of the Montreal effluent outfall, especially in sediments. Likewise, the slight, but not significant, bioconcentration of certain alkylphenol polyethoxylates (AP(n)EO) in the mussels was more noticeable at the downstream site than at the upstream site.

A multispecies approach for monitoring persistent toxic substances in the Gulf of Gdansk (Baltic sea).

Bivalve mussels are usually used for biomonitoring persistent toxic substances (PTS) in coastal ecosystems. Nevertheless, these organisms, which live attached on hard substrates, can be found along the sandy coasts only on human manufactured products. In this work different species collected in the Gulf of Gdansk were compared to evaluate their suitability for monitoring PTS pollution at a local scale. The clam Mya arenaria seems to represent an excellent indicator of sediment pollution, mainly for organotin compounds which are selectively bioaccumulated. Organochlorine compounds are bioaccumulated in the different species mainly in function of their lipid body burden. Habitat conditions (salinity, substrate, pollution), however, strongly limited the occurrence of different species in the sampling sites; the most ubiquitous species, the common shrimp Crangon crangon, resulted therefore the most suitable to be used for the comparison of PTS pollution in this aquatic environment. Although the blue mussel (Mytilus trossulus) was confirmed to be a very useful sentinel species to compare pollution level inside and outside the Gulf of Gdansk, we recommend the use of other species to give a more detailed picture of the pollution situation in coastal areas.

Some chemical contaminant of surface sediments at the Baltic Sea coastal region with special emphasis on androgenic and anti-androgenic compounds.

Androgenic and anti-androgenic compounds including p,p'-DDE, Diuron, Linuron, Fenarimol, Vinclozolin, 1-(3,4-dichlorophenyl) urea (DCPU), 1-(3,4-dichlorophenyl)-3-methylurea, (DCPMU), tributyltin (TBT) and triphenyltin (TPT) and their metabolites (DBT, MBT, DPT, MPT) as well as metallic elements (Ni, Cu, Zn, As, Cd, Pb, Co, Tl, Cr, Fe, Mn, Al, K, Mg, Na, Ca, Ba, Ti, Sn), PAHs (16 indicator compounds), DDTs and PCBs have been quantified in top layer (0-10 cm) of up to 37 surface sediment samples collected from several sites in costal zone of the Gulf of Gdansk, an inland freshwater area of Brdyujscie in Poland and the tidal flats of the Norderney Island, Wadden Sea in 2002-2003. These sites differed in the degree of anthropogenic activities, including chemical pollution and related impact on biota. Especially in sediments near shipyards, ship repair facilities, harbours, other industrial activities or close to municipal sewage treatment plant outlets butyltins, PAHs and some metallic elements were found at high concentrations. Diuron, Linuron and DCPMU were detected at a few sites, Fenarimol only once, while Vinclozolin and DCPU were not detected. DDT concentrations in the sediments from the Gdansk and Gdynia region of the Gulf show a stepwise decrease following the ban for production and use, while diffusion of PCBs at some industrial sites seems to continue. Elevated PAH concentrations in sediments seem to be mainly due to pyrogenic and less to mixed pyrogenic and petrogenic sources, while for a few sites rather petrogenic sources dominated. The reference sites in the Norderney Island, Wadden Sea showed similar or slightly higher loads of DDTs, BTs, PAHs, PCBs and metallic elements when compared to sediments from the least contaminated sites in the coastal Gulf of Gdansk area, while phenyltins were not detected at both spatially distant European areas.