Einsatz eines Immunoassays als Screening-Verfahren für die Untersuchung von Grundwasserproben auf Mecoprop

Application of an Enzyme Immunoassay for the Detection of Mecoprop in a Ground water Screening Procedure A competitive solid-phase enzyme immunoassay based on rabbit polyclonal antibodies was tested for its reliability. This assay, used for the detection of the phenoxycarboxylic acid mecoprop [2-(4-chloro-2-methylphenoxy-)propionic acid], has already been characterized in terms of middle of the test (0.36 μg/L at 50% B/B₀) and detection limit (0.06 μg/L at 80% B/B₀) by Weber (Acta hydrochim. hydrobiol. 22, 53–59 (1994)). For the reliability test, a small number from a batch of one hundred groundwater samples were spiked. The analyst was neither aware of which nor of how many samples were spiked. The spiked samples were all detected as positive. There were no false negative results. From these findings, values of 1 were obtained for the diagnostic sensitivity and for the diagnostic specificity. Based on these results, the mecoprop contents of 228 groundwater samples from various waterlevel gauges of a test field were determined. The test site had previously been treated repeatedly with this herbicide. Confirmation of the enzyme immunoassay results was carried out by GC/MS. This showed that the enzyme immunoassay gave one false positive result only. All other results from immunoassay and GC/MS showed good correlation.     

Entwicklung eines Enzymimmunoassays zum Nachweis des Herbizids Mecoprop in Trink- und Grundwasser auf der Basis von Antikörpern aus den Eiern von Legehennen

Development of an Enzyme Immunoassay for the Detection of Mecoprop in Drinking Water and Groundwater Based on Antibodies Raised in Chicken Egg Yolk Antibodies against mecoprop were isolated from egg yolk of immunized hens and were used for the development of an enzyme-linked immunosorbent assay. We tested assay parameters (pH and concentration of buffer, incubation temperature, kind of enzyme tracer) to optimize the standard curve for mecoprop. By decreasing the pH, the detection limit was reached at a concentration of 0.35 μg/L mecoprop. The concentration for 50% inhibition (50% B/B₀) was 2.8 μg/L. Dichlorprop and the methyl esters of both mecoprop and dichlorprop showed high crossreactivity (165%, 400% and 233%). Antibodies against mecoprop separated from egg yolk were compared to antibodies raised in rabbit for both sensitivity and specificity. Chicken immunoglobulins were found to be less sensitive and specific than the mammalian IgG's.     

Die gaschromatographische Bestimmung von Diethylentriaminpentaessigsäure in Oberflächenwasser

Gaschromatographic Determination of Diethylenetriaminepentaacetic Acid in Surface Water. A method for the determination of diethylenetriaminepentaacetic acid (DTPA) is reported. The determination is based on the preconcentration on a strong base anion exchange resin, esterification with n-propanol and following gaschromatographic separation using a N-selective detector. The sensitivity of the method, including enrichment, is 1 μg/L in river water. The identity of DTPA in river water was proved by mass spectrography. The method can be used also for the quantitative determination of nitrilotriacetic acid (NTA), ethylenediaminetetraacetic acid (EDTA), cyclohexanediaminetetraacetic acid (CDTA) and ethyleneglycolbis-(2-aminoethylether)-N,N,N′,N′-tetraacetic acid (EGTA). The method was used to examine river water. An investigation along the river Rhine, taking grab samples from January and february 1992 showed that DTPA was present in some sampling places in similiar concentrations as NTA and EDTA.     

Improvement of a Monoclonal Antibody-based Immunoassay for the Determination of Terbutryn Verbesserung eines Immunassays mit monoklonalen Antikörpern zur Bestimmung von Terbutryn

The performance of an existing enzyme immunoassay (EIA) with monoclonal antibodies (mAb) for the determination of terbutryn was improved by the application of a new enzyme tracer. For this purpose the triazine derivative 6-chloro-2-(tert-butylamino)-1,3,5-triazine-4-(6-aminohexane carboxylic acid) was coupled to horseradish peroxidase (HRP). The competitive EIA on microwell plates made it possible to determine terbutryn in the range from 0.05 to 1 μg/L with a 50% B/B₀ value of the test at 0.2 μg/L. The application of the EIA to determine terbutryn in spiked surface waters provided good recoveries of terbutryn without matrix effects.     

Nachweis von Industriechemikalien (HPS,BPS und SPS) im Oberflächenwasser

Occurrence of Industrial Chemicals (HPS, BPS, and SPS) in Surface Water The paper gives the results of water examinations for different phenylsulfonamides. Random samples taken every month between May 1999 and August 2000 from surface water out of the river Rhine (kilometer 838), the river Ruhr (Mülheim Styrum) and the river Emscher (Oberhausen center) were tested for the corrosion inhibiting agent 6‐[methyl(phenylsulfonyl)amino]‐hexanoic acid (HPS) as well as its metabolites 4‐[methyl‐(phenylsulfonyl)amino]‐butanoic acid (BPS) and sarkosin‐N‐(phenylsulfonyl) (SPS). Furthermore, the sewage plant effluents of two municipal wastewater treatment plants from the rural area were also included in the monitoring program. The analytical method includes solid‐phase extraction (SPE), a derivatization step as well as gas chromatography mass spectrometry (GC‐MS). SPS is regularly found in all investigated surface waters, but only occasionally in the effluents of the two rural sewage plants. The median values for SPS amount to 0.09 μg/L in the river Rhine, 0.60 μg/L in the river Ruhr, and 0.70 μg/L in the river Emscher. BPS can only be found in the river Ruhr (median value: 0.08 μg/L) and in the river Emscher (median value: 0.41 μg/L). HPS was regularly found in a surface water for the first time. This substance can be detected in the Emscher through the whole measurement period. The median value for HPS amounts to 1.78 μg/L. Aditionally, the validation characteristics of an alternative analytical method including solid‐phase microextraction (SPME) is worked out. The fully automated process includes an on‐fiber methylation step and the GC‐MS. The repeatability standard deviation of the process amounts to RSD < 12%. Detection limits between 0.07 and 0.70 μg/L are achieved.     

Abschätzung und Beeinflussbarkeit der Arsenmobilität in kontaminierten Böden

Assessment and Modification of Arsenic Mobility in Contaminated Soil Arsenic concentration in the seepage of contaminated soils of an old tannery site is assessed using batch and column experiments. The effect of reducing conditions, pH, and ionic strength is also investigated. The iron oxide rich subsoil (C‐horizon) is the main source of groundwater pollution with arsenic. In this horizon, mobilization can increase as a result of reducing conditions upon periodical water saturation. Therefore, the potentially mobile arsenic is determined by a reductive dissolution of the poorly crystalline iron oxide fraction using 0.1 M ascorbic acid. Arsenic concentration can be reduced from 100 μg/L to below 20 μg/L by an increase of ionic strength (e.g. by a 0.01 M CaCl₂ solution). Arsenic contaminated soils should be limed regularly in order to maintain the highest possible calcium concentration in the soil solution.     

Bestimmung von Umweltqualitätsnormen für potenziell gewässerrelevante Stoffe

With the Water Framework Directive (WFD) the European Commission (EC) is requested to define environmental quality standards for pollutants to protect aquatic life in surface waters. Quality standards have to be derived by a scientific risk assessment and should not be exceeded. The recommended quality standards are mainly based on long term toxicological tests with algae, crustaceans and fish. The lowest effect concentration is divided by an assessment factor between 10 and 1000, which considers the data quality and quantity. In this work funded by the German Länder Working Party on Water (Länderarbeitsgemeinschaft Wasser – LAWA) environmental quality standards for 40 considered relevant substances were derived for aquatic ecosystems. The data sheets cover the identification of the compounds, their behaviour in the environment (physical‐chemical properties, biotic and abiotic degradation, sorption, bioaccumulation), information about mode of action, uses, analytical determination and available quality criteria. Ecotoxicological effect concentrations for bacteria, algae, protozoa, aquatic plants, crustaceans, fish, amphibians, insects and molluscs are listed. For 17 of the 40 hazardous substances investigated, quality objectives above 1 μg/L were derived. For 12 substances the values were between 0.1 μg/L and 1 μg/L and for 5 substances lower than 0.1 μg/L. Incomplete ecotoxicological data sets of the remaining compounds do not allow the derivation of quality standards. Especially for drugs further ecotoxicological test results are needed.     

Occurrence and Distribution of Organic Contaminants in the Aquatic System in Berlin. Part I: Drug Residues and other Polar Contaminants in Berlin Surface and Groundwater

Several polar contaminants were found in screening analyses of 30 representative surface water samples collected from rivers, lakes, and canals in Berlin. Residues of pharmaceuticals and N-(phenylsulfonyl)-sarcosine originating from various sewage treatment plants effluents were found at concentrations up to the μg/L-level in the surface water, whereas the concentrations of polar pesticides such as dichlorprop and mecoprop were always below 0.1 μg/L. The pharmaceuticals most frequently detected in the surface water samples include clofibric acid, diclofenac, ibuprofen, propiphenazone, and two other drug metabolites. Additional investigations of groundwater wells of a drinking water plant have shown that polar contaminants such as drug residues or N-(phenylsulfonyl)-sarcosine easily leach through the subsoil into the groundwater aquifers when contaminated surface water is used for groundwater recharge in drinking water production.     

Dünnschichtchromatographischer Nachweis von Mineralöl in Brunnenwasser

A tabular survey is given from literature concerning seven extraction techniques, 14 thin-layer chromatographic detection techniques and 15 spray reagents. From the possibilities represented in this way, a newly composed method is described which can be carried out in any routine laboratory within one hour and has proved successful in situations of average of the drinking water supply. 1 l water is extracted with 25 ml CCl₄, the extract is concentrated in the vacuum to 1 ml and chromatographed with n-hexane on a prefabricated foil. Detection is carried out with 10% methanolic molybdophosphoric acid. The detection limit is at 0.5 μg.     

AOX‐Bildung bei der sonolytischen Oxidation von Salicylsäure in Gegenwart von Chlorid

AOX‐formation by the Sonochemical Treatment of Salicylic Acid in Presence of Chloride Ultrasound shows great potential for improving water, wastewater and sludge treatment processes. However, a number of questions exist: for example the influence of suspended solids or salts. In this paper the influence of chloride in view of AOX‐formation should be investigated. As organic model compound salicylic acid (0.05…1 mmol/L) was used. Formation of AOX (adsorbable organically bound halogens) was measured in dependence upon chloride concentrations (1.4…141 mmol/L) and pH 2 and 9. Ultrasound irradiation was performed in glass reactor (500 mL) at 206 kHz and 353 kHz by continuous bubbling argon:oxgen (4:1) at a flow rate of 1 L/min into the solutions. The elimination of salicylic acid and the formation of hydrogen peroxide were not influenced by the presence of chloride (up to 56 mmol/L). The rate of salicylic acid elimination and the formation rate of hydrogen peroxide are at concentrations of mmol/(L min) levels but those of AOX formation of μmol/(L min) levels. The reactions leading to chlorinated by‐products can be seen as minor reaction paths. The yield of AOX depending on frequency, pH and chloride concentration lies between 10 μg/L and 900 μg/L. In acidic medium they were twice as high as in basic medium. Below the ratio chloride : salicylic acid of 2 mol/mol, AOX could not be identified.     

Präzipitation von Aluminium und Phosphat in Gewässern unter dem Einfluss von Versauerung

Precipitation of Aluminium and Phosphate Affected by Acidification Acidified waters often show elevated concentrations of Al (with up to 6 mg L^–1 being not unusual). A pH increase resulting e.g. from mixing with non‐acidified water or from biological activities may be linked with Al precipitation. Up to now, this phenomenon was described for acid mine drainages. This investigation focuses on a whitish precipitate naturally formed in a brook of an atmospherically acidified catchment in the Ore Mountains, Germany. Based on infrared spectra the precipitate was identified as an Al‐hydroxosulfate with crystal water. A simulation of natural conditions in the laboratory showed that Al precipitated only if sulfate or phosphate ions were added to the solution. In the case of sulfate being added, the infrared spectrum of the precipitate was similar to the natural precipitate. ²⁷Al NMR spectroscopy revealed tetrahedrally coordinated Al in some precipitates which evidences the participation of the tridecameric [Al₁₃O₄(OH)₂₄(H₂O)₁₂]⁷⁺ cation beside other polymeric Al cations. Precipitation experiments subjected to the given conditions showed that the phosphate elimination from solution with Al was much higher than with Fe. With Al and Fe added together, the P elimination rate was likewise high, and phosphate was bound onto Al in the precipitate. This was demonstrated by SEM‐EDX spectroscopy. Based on these results we present a possible reaction mechanism. The precipitation of Al together with P allows a significant retention of both elements in sediments because in contrast to Fe, Al immobilizes phosphate even under anoxic conditions.     

Bildung bromhaltiger Desinfektionsnebenprodukte nach Chlorung von Bodenseewasser in Abhängigkeit von Aufbereitungsschritten und der Bromidkonzentration

Formation of Brominated Disinfection By-products after Chlorination of Water from Lake Constance Depending on Treatment Steps and Bromide Concentration The formation of halogenated disinfection by-products (DNP) (trihalomethanes (THM), haloacetic acids (HES), haloacetonitriles (HAN), haloketones (HKE) and chloropicrin (TCNM)) was studied after chlorination (0.5 mg/L) of raw water and drinking water from Lake Constance. The objectives of this study were to investigate the effect of bromide ion on the distribution of DNP-species resulting from chlorination and to draw up a bromide mass-balance. The treatment of water from Lake Constance with ozone decreased the formation of DNP about 35% and of total organic halogens (AOX) about 45%. This decline in DNP-formation is mainly due to the formation of chloroform, dichloro- and trichloroacetic acid. Though there is only a small amount of bromide (6.2 μg/L) in water from Lake Constance, about 32% of THM, 16% of HES, and 30% of HAN contained bromine after chlorination of treated water. The addition of bromide ion (up to 1 mg/L) shifted the DNP-composition to more brominated species. The distribution of the DNP-species depends on the initial molar bromide-to-chlorine ratio. A mathematic model showed the same pattern of DNP-species depending on the initial molar bromide-to-chlorine ratio that was observed experimentally in microstrained raw water and drinking water from Lake Constance. After chlorination of microstrained raw water, 27% of the bromide were incorporated in known DNP, 34% in unknown substances, and about 39% of bromide were not converted to organic bromine. After chlorination of treated water, only 16% of the bromide were incorporated in unknown substances, whereas 34% were found in known DNP. About 50% were found as bromide again. After addition of bromide (up to 25 μg/L) to ozonated and treated water from Lake Constance, between 60 and 72% of the bromide that had reacted after chlorination were found in analyzed low-molecular DNP.     

Occurrence and Distribution of Organic Contaminants in the Aquatic System in Berlin.Part II: Substituted Phenols in Berlin Surface Water

In a surface water screening, 30 representative surface water samples collected from rivers, canals, and lakes in Berlin were investigated for the presence of 22 substituted phenols. The phenols selected include the 11 phenols considered as “priority pollutants” by the US Environmental Protection Agency (US-EPA). Surface water samples were extracted applying solid-phase extraction with styrenedivinylbenzene adsorbent. The recoveries, determined in spiking experiments, were between 80 % and 103 %. After derivatization with N-(tert-butyldimethylsilyl)-N-methyl-trifluoroacetamide (MTBSTFA) the samples were analyzed by capillary gas chromatography-mass spectrometry (GC-MS). Phenol, cresols, 2-ethylphenol, 2-chlorophenol, 4-chloro-3-methylphenol, pentachlorophenol, 2-nitrophenol, and 4-nitrophenol were detected in the surface water samples at concentrations between 0.02 μg/L and 7.8 μg/L, respectively. The distribution of these residues in the Berlin surface waters showed that the phenolic residues, with the exception of pentachlorophenol and 2-ethylphenol, do not originate primarily from municipal sewage treatment plants discharges. Some of the phenols are formed naturally or occur as ubiquitous anthropogenic contaminants in the aquatic system.     

Fate of Point‐Source Pesticide Spill in Clayey Till after 15 Years

This study investigated the development of pesticide pollution two, three, and 17 years after spills of the herbicides dichlorprop, mecoprop (MCPP), MCPA, 2,4‐D (phenoxy acids), simazine, and terbutylazine (triazines) in a former orchard machinery service yard. The spills had occurred over several decades on a 23‐m thick, mainly anaerobic fractured clayey till aquitard. Angled monitoring wells were installed in the aquitard 3 years after the spills ceased in 1989. In 1993, monitoring revealed that high groundwater concentrations of dichlorprop (677 µg/L) and MCPP (139 µg/L) were accumulated as a zone of maximum pollution in anaerobic and largely immobile pore water at 5 to 6 m depth in the aquitard profile. In contrast, 2,4‐D was determined in only one water sample, and MCPA and simazine and terbutylazine were determined only in low concentrations (below10 µg/L), although these pesticides had been handled at the site in greater amounts than dichlorprop and MCPP according to detailed historic information obtained for the site. Repeated monitoring in the same wells after a further 14 years in 2007 revealed that no identifiable degradation of MCPP had occurred, while dichlorprop had degraded by 75% to 80% (estimated half‐life of approximately 5 years). Furthermore, degradation products related to the phenoxy acids had accumulated, especially 4‐CPP with a maximum concentration of 218 µg/L. In the same zone, MCPA and simazine had almost disappeared. As the pollution was mainly accumulated in largely immobile pore water of the aquitard clayey matrix, and the groundwater recharge was low (30 to 60 mm/year), only minor vertical displacement of the zone with maximum pollution zone had occurred during the 15 years of monitoring. However, concentrations of dichlorprop (0.01 to 0.02 µg/L), MCPP (0.1 to 0.2 µg/L), and 4‐CPP (0.6 to 0.7 µg/L) had spread along textural heterogeneities in the aquitard into the underlying sandy aquifer at 23‐m depth.     

Polarographische Bestimmung von Schwermetallen im Klärschlamm

The differential-pulse polarography (DPP) and the stripping voltametry (SV) are investigated in detail with respect to their suitability for the quantitative detection of individual traces of heavy metals in sewage sludge. The results are checked on the basis of AAS analyses and by the standard-addition method. From the hydrochloric-acid extracts of fused sludge samples down to 1 μg/l can be detected by the SV, whereas the DPP reaches a sensitivity of 100 μg/l. The following basic electrolytes are used: Zn: 2 … 3 mol/l H₃PO₄; Cu: 0.4 mol/l K₂CO₃, 0.2 mol/l Na-K-tartrate, 0.1 mol/l HCl; Ni: 1 mol/l NH₄OH, 1 mol/l NH₄Cl, 25 ml/l triethylamine; Pb and Cd: 0.1 … 0.2 mol/l HCl.     

Development of an Enzyme Immunoassay for the Analysis of the Atrazine Metabolite Hydroxyatrazine Entwicklung eines Enzymimmunoassays zur Analyse des Atrazin-Metaboliten Hydroxyatrazin

A highly sensitive and specific enzyme immunoassay (EIA) is described for the detection of the atrazine metabolite hydroxyatrazine. Polyclonal antibodies were raised in rabbits by immunization with a hapten-bovine serum albumin (BSA) conjugate containing 8 hapten residues per molecule of BSA. An EIA with a horseradish peroxidase (HRP) hapten tracer was optimized in microtitre plates. A concentration of 50% B/B₀ was found at 0.10 μg/L for hydroxyatrazine. A limit of determination for hydroxyatrazine was reached at approximately 0.01 μg/L, i.e. well below the maximum concentration permitted by the EU guidelines for drinking water and the drinking water ordinance of the FRG. The assay did not require concentration or clean-up steps for drinking water or ground water samples. Validation experiments confirmed a good accuracy and precision. Hydroxyatrazine is reported to be the main atrazine metabolite found in soil samples. As organic solvents are usually employed for soil extraction, the influence of methanol as representative organic solvent on the assay was examined. Up to a concentration of 5% (v/v) methanol, the organic solvent did not affect the assay.     

Zum Auftreten von leichtflüchtigen Halogenkohlenwasserstoffen im Badewasser

110 samples from two swimming-pools were gas-chromatographically investigated for halogenated hydrocarbons. In spite of varying chlorine concentrations between 0.1 and 2.5 mg/l Cl₂ the bacteriological results were unobjectionable. The total of the haloforms was between 1.2 and 182 μg/l, with the average value of 90 μg/l. Besides CHCl₃, CHBrCl₂, CHBr₂Cl and CCl₃NO₂ there were observed other, still unidentified compounds in the chlorinated bathing water which clearly are successive products of disinfection, too. The water treatment by flocculation and filtration does not bring about any elimination of trihalomethanes. Of special importance is the occurrence of trichloronitromethane in concentrations of 0.13 … 1.2 μg/l, whose LC₅₀ in the Daphnia test is 189 μg/l, at a threshold concentration of 160 μg/l. To limit the pollution of water and air to a minimum, specific conditions must be adhered to in the operation of indoor swimming-pools.     

Bestimmung von Atrazin,Terbutylazin und ihrer Desethyl- sowie Hydroxymetaboliten in Boden mittels SFE und HPLC/UV-DAD

Determination of Atrazine, Terbutylazine and their Deethyl- and Hydroxymetabolites in Soil by SFE and HPLC/UV-DAD The supercritical fluid extraction (SFE) (CO₂ and CH₃OH as modifier) of the herbicides atrazine and terbutylazine as well as their deethyl- and hydroxymetabolites from soil was investigated. The analytes were determined after extraction by HPLC/UV-DAD using a C18 column. The limits of detection of the chromatographic step were between 0.01 μg/mL and 0.07 μg/mL, the relative standard deviations between 0.8% and 1.4%. With the aid of a control chart, the stability of the chromatographic system was evaluated. For extraction, soil was spiked with 5 μg/g for each component. For SFE, CO₂ and methanol as modifier were used, and a pressure program was applied. Beside SFE, Soxhlet extraction with methanol and a solid-liquid extraction with acetone/water by shaking were carried out. For chlorine-containing triazines, the recovery rates were comparable for all extraction methods. The following recovery rates for SFE were obtained: deethylterbutylazine 77.4%, terbutylazine 80.2%, deethylatrazine 87.4%, atrazine 92.6%. However, the recovery rates for the hydroxymetabolites (4.1% for hydroxyatrazine, 21% for hydroxyterbutylazine) were not satisfying compared with the “classical” methods of extraction (above 50%). The limits of detection for the total process (SFE and HPLC/UV-DAD) estimated by the signal to noise ratio amounted 0.08 μg/g soil for chlorine-containing derivates, for hydroxyatrazine 2…3 μg/g, and for hydroxyterbutylazine 0.8 μ/g.     

Removal of Various Phenoxyalkanoic Acid Herbicides from Water by Organo‐clays

Dodecylammonium bentonite (DB) and dodecylammonium sepiolite (DS) were used as sorbents for phenoxyalkanoic acid herbicides 2,4‐D ((2,4‐dichlorophenoxy)acetic acid), 2,4‐DP ((RS)‐2‐(2,4‐dichlorophenoxy)propionic acid), 2,4‐DB (4‐(2,4‐dichlorophenoxy)butyric acid), 2,4,5‐T ((2,4,5‐trichlorophenoxy)acetic acid), and MCPA ((4‐chloro‐2‐methylphenoxy)acetic acid). Langmuir, Freundlich, and the linear Henry’s Law isotherm adsorption parameters were calculated from the adsorption isotherms. Langmuir equation showed poor fit for both adsorbents. According to the evaluation using the Freundlich equation, the DS sample showed much higher and stronger sorption capacity than DB. Similar behaviour was also observed in the case of the linear Henry’s Law isotherm. The adsorption of the herbicides on both DB and DS decreased in the order of 2,4‐DB > 2,4,5‐T > 2,4‐DP > 2,4‐D > MCPA.     

Bestimmung von Pflanzenbehandlungsmitteln in Wasser durch Online-Fest-Flüssig-Extraktion und HPLC-DAD

Determination of Pesticides in Water by On-line Solid-phase Extraction and HPLC-DAD This paper describes an HPLC on-line procedure for the determination of 29 relevant pesticides in water by diode-array detection using a compact HPLC-system with integrated column-switching. The sample – 5 mL – is injected with a large volume injection system on a precolumn cartridge (RP-C18). Enriched compounds are eluted in frontflush mode by columnswitching to a microbore column and separated with a gradient water/acetonitrile. During desorption the precolumn cartridge is heated at 80°C resulting in a comparable efficiency to the separation with direct injection. The procedure is fully automated for continuous monitoring of drinking-, ground-, and surface water with low contamination levels. Sample preparation is limited to a filtration step with an anion-exchange membrane filter to reduce natural contaminations e.g. humic acids simultaneously. For a concentration of 50 ng/L, the signal to noise ratio varies from 3:1 (Ethidimuron) up to 25:1 (Chloridazon). The standard deviations compare to the standardized HPLC-method DIN EN ISO 11369; the recovery rates are 100% and reproducible. The method was successfully tested on ruggedness with several sequences from a monitoring program. The precolumn cartridge needs replacement after 100 sample injections. Finally, this method was coupled with a benchtop mass-spectrometer (electrospray mode) without changing the chromatographic conditions. With selected ion monitoring (SIM) selectivity and detection sensitivity could be improved considerably compared to DAD-detection.