Stability study and determination of benzene- and naphthalenesulfonates following an on-line solid-phase extraction method using the new programmable field extraction system

Seven benzene- and naphthalenesulfonates (3-nitrobenzenesulfonate, 4-methylbenzenesulfonate, 1-hydroxy-4-naphthalenesulfonate, 1-amino-7-naphthalenesulfonate, 4-chlorobenzenesulfonate, 1-naphthalenesulfonate and 2-naphthalenesulfonate) were studied. A rapid method for quantifying aromatic sulfonated compounds from waste water samples was developed. This method consists in on-line in-field sampling and monitoring based on ion-pair solid-phase extraction with PLRP-S sorbent, using the new programmable field extraction system and ion-pair liquid chromatography with UV diode-array and electrospray mass spectrometry. Limits of detection for the studied compounds, using the SIM acquisition mode, ranged from 0.01 to 0.33 ng ml(-1). The influence of the aqueous matrix on the on-line SPE was checked by spiking ground and waste waters. Recoveries varied from 70 to 99% when 10 ml of water sample were enriched. The method was applied to the analysis of some environmental sewage samples. This study confirmed that high concentration levels of aromatic sulfonated compounds can be found in sewage samples. In addition, the stability of the seven studied sulfonated benzene and naphthalene compounds was investigated using on-line polymeric SPE pre-columns, based on the styrene-divinylbenzene polymer PLRP-S. Different storage conditions were tested to carry out the stability survey, which included storage at room temperature, at 4 degrees C and at -20 degrees C, during a period of up to 2 weeks. This study showed that the stability of aromatic sulfonic acids on disposable on-line SPE polymeric pre-columns is related to temperature and that the target compounds are more stable at lower temperatures.     

Simultaneous determination of aliphatic and aromatic amines in water and sediment samples by ion-pair extraction and gas chromatography-mass spectrometry

A gas chromatography-mass spectrometry (GC-MS) method has been proposed for the determination of aliphatic and aromatic amines in a variety of environmental samples including wastewater, river water, sea water and sediment samples. The method includes ion-pair extraction with bis-2-ethylhexylphosphate (BEHPA), derivatisation of compounds with isobutyl chloroformate (IBCF) and their GC-MS analysis. Aliphatic and aromatic amines were isolated from aqueous samples using BEHPA as ion-pair reagent and derivatised with IBCF for their chromatographic analysis. Solid-liquid extraction of aliphatic and aromatic amines in sediment samples were performed in Soxhlet apparatus with acidic MeOH and ion-pair extraction with BEHPA were carried out for the isolation of amines followed by derivatisation with IBCF. Aliphatic and aromatic amines were then analysed with GC-MS in both electron impact (EI) and positive and negative ion chemical ionisation (PNICI) mode as their isobutyloxycarbonyl (isoBOC) derivatives. The obtained recoveries ranged from 81.0 to 98.0% and the precision of this method, as indicated by the relative standard deviations (RSDs) was within the range of 0.5 and 4.3%. The detection limits obtained from calculations by using GC-MS results based on S/N = 3 were within the range from 0.07 to 0.50 ng/l.     

Determination of nitrite as pentafluorobenzyl derivative by RP-HPLC and UV detection with and without ion-pair extraction

Summary A reversed-phase high-performance liquid chromatographic (RP-HPLC) method with UV detection (270 nm) for the determination of nitrite as its pentafluorobenzyl derivative with and without ion-pair extraction is described. Ion-pair extraction of nitrite from aqueous solutions was performed by using a 1 mol/l solution of the liquid ion exchanger methyltrioctylammonium chloride in toluene. The residue of the ion-pair extraction or an aliquot of an aqueous nitrite solution or of a biological fluid (100 l) were treated with 400 l of acetone and 10 l of pentafluorobenzyl bromide. Nitrite was converted into its pentafluorobenzyl derivative by heating at 50°C for 90 min. After evaporation of acetone the aqueous phases were diluted with 100 to 400 l of methanol, and up to 100 l were injected into the RP-HPLC system. The method allows accurate analysis of nitrite in the presence of nitrate directly in aqueous solutions and biological fluids in concentrations down to 2.0 mg/l. The method is also applicable to the determination of nitrate following its reduction to nitrite by cadmium.     

Microwave Assisted Extraction of Polycyclic Aromatic Hydrocarbons and their Determination by Gas Chromatography–Mass Spectrometry: Validation of the Method and Application to Marine Sediments

Microwave-assisted extraction of sixteen polycyclic aromatic hydrocarbons and their gas chromatographic mass spectrometric detection are presented herein. An efficient extraction was achieved in 15 minutes using 10 mL of 1:1 n-hexane-acetone while a clean-up step was developed studying the elution curves on solid phase extraction silica cartridges. The analytical method was optimized and validated using a certified reference marine sediment; satisfactory figures of merit were obtained with limits of detection in the range 0.001–0.004 µg/g, precision within 6%, and good linearity (regression coefficients generally higher than 0.998, in the concentration range 0.010–1.000 µg/mL). The developed method was successfully applied to the determination of polycyclic aromatic hydrocarbons in real marine sediments collected in two coastal areas of Italy exposed to different anthropic impact: three tourist sites of Liguria and the Venetian Lagoon. The total concentration of the analytes in the samples was in the range 1.027–3.827 µg/g and the use of common markers suggested their probable pyrolytic origin.     

Fully automated high-performance liquid chromatographic assay for the analysis of free catecholamines in urine

A totally automated and reliable high-performance liquid chromatographic method is described for the routine determination of free catecholamines (norepinephrine, epinephrine and dopamine) in urine. The catecholamines were isolated from urine samples using small alumina columns. A standard automated method for pH adjustment of urine before the extraction step has been developed. The extraction was performed on an ASPEC (Automatic Sample Preparation with Extraction Columns, Gilson). The eluate was collected in a separate tube and then automatically injected into the chromatographic column. The catecholamines were separated by reversed-phase ion-pair liquid chromatography and quantified by fluorescence detection. No manual intervention was required during the extraction and separation procedure. One sample may be run every 15 min, ca. 96 samples in 24 h. Analytical recoveries for all three catecholamines are 63-87%, and the detection limits are 0.01, 0.01, and 0.03 microM for norepinephrine, epinephrine and dopamine, respectively, which is highly satisfactory for urine. Day-to-day coefficients of variation were less than 10%.     

Gas-chromatographische und dünnschicht-chromatographische Bestimmung von Eulan® WA neu in Abwässern

Two methods for the determination of the active ingredient of Eulan WA neu in textile industry waste waters are described:

1. Gas chromatographic method. The waste water is acidified and extracted with toluene. The active ingredient is determined in the extract with ECD (⁶³Ni)-indication by external standards.
2. Thin-layer chromatographic method. After having splitted the active ingredient—directly in the waste water or after extraction with toluene—the resulting aromatic amines are separated by TLC. The spots are coloured by diazotizing and coupling with 1-N-naphthylethylene-diamine. Evaluation is made by comparison with standards. Taking 500 ml of waste water per analysis the detection limit is 0.05 ppm for both the methods.

     

Determination of benzene metabolites in urine of mice by solid-phase extraction and high-performance liquid chromatography.

A method was developed for quantitative measurement of trans,trans-muconic acid, catechol, hydroquinone and phenol in urine. Hydrolysis of esterified and glucuronized phenolic compounds was effected by specific enzymes. The hydrolysed mixture was purified and separated by solid-phase extraction with an anion exchanger, followed by extraction with diethyl ether. By using a clean-up procedure the natural background from mouse urine could be reduced, so that the detection limit of the metabolites was in the range 3-60 mg/l. Optimization of the chromatographic conditions resulted in a short high-performance liquid chromatography analysis time. Phenol had the longest retention time of about 10 min. The clean-up procedure could also be used for phenylmercapturic acid, an additional benzene metabolite, but for sensitive high-performance liquid chromatographic detection of phenylmercapturic acid other conditions are necessary.     

Characterization of an avidin-bonded column for direct injection in reversed-phase high-performance liquid chromatography

An automatic method for the determination of metabolites of Ropivacaine in urine was set up. It utilizes supported liquid membrane extraction for sample clean-up and enrichment, followed by ion-pair chromatography determination using UV detection. The extraction was very selective with no observed interfering compounds from the urine matrix, permitting simple isocratic chromatographic analysis. The detection limits for spiked urine samples were 2–18 nM for the different compounds. The repeatability was 1–3% (RSD) with an internal standard that was also extracted, and about twice without this standard. A throughput of 3.3 samples per hour was achieved and the liquid membrane was stable for more than a week.     

Estimation of tetrahydro, dihydro and fully oxidised pterins by high-performance liquid chromatography using sequential electrochemical and fluorometric detection

A method is described for the separation and detection of tetrahydro, dihydro and fully oxidised pterins in a single chromatographic run using ion-pair reversed-phase high-performance liquid chromatography. Tetrahydropterins are detected by electrochemical oxidation, dihydropterins by fluorescence following post-column electrochemical oxidation and the fully oxidised pterins by their natural fluorescence. The post-column electrochemical conversion of the non-fluorescent dihydropterins to fluorescent compounds is proportional to the amount injected over three orders of magnitude. Because of the relative selectivity of the fluorescence detection and the low potential required to oxidise the tetrahydropterins, all the oxidation species of the pterins may be measured in biological samples with minimal sample clean-up.     

Comparison of two extraction methods for the analysis of per- and polyfluorinated chemicals in digested sewage sludge

A rapid and reliable analytical method, based on ion-pair extraction, clean-up on Envicarb cartridge and detection by liquid chromatography–tandem mass spectrometry (LC–MS/MS), was developed for determination of 17 per- and polyfluorinated chemicals (PFCs) in digested sewage sludge. Envicarb cartridge and six labeled internal standards were selected for the elimination/reduction and correction of matrix effects, respectively. As a result, the matrix effect for perfluorooctane sulfonamides (FOSAs) and perfluorocarboxylic acids (PFCAs) with carbon chain length from C6 to C14 was lowered to a range of −14% to +28%. However, the matrix effect for other analytes was still great mainly due to the absence of appropriate internal standard. Mean recoveries of the target analytes based on matrix spikes, at different spike levels (10–300 ng/g), ranged from 70% to 169%. Relative standard deviations (RSDs) were in the range of 2–20% at different spike levels. The limit of quantification (LOQ) ranged between 0.6 and 30 ng/g. The method was successfully applied to several sewage sludge samples from wastewater treatment plants nearby Zürich, Switzerland. In addition, by comparing the accuracy and precision of ion-pair extraction method and methanol extraction method, we further demonstrated that the ion-pair extraction method can be used for the analysis of PFCs in sludge samples. To our knowledge, this is the first study to extract the PFCs in sewage sludge with ion-pair method and to find unsaturated fluorotelomer carboxylic acids (FTUCAs) in sewage sludge.     

Determination of fluazifop-butyl and fluazifop with the use of disposable solid phase extraction columns for selective clean-up and concentration of Soxhlet soil extracts

Summary An analytical procedure based on the solid phase extraction technology has been developed for the clean-up and concentration of Soxhlet soil extracts containing fluazifop-butyl and fluazifop by the use of a phenyl phase cartridge. No liquid-liquid partition has been used; thus the consumption of organic solvents was limited and the use of chlorinated solvents could be avoided. Quantification has been performed by ion-pair HPLC. Despite the large difference in polarity the recoveries of both the compounds from spiked soil samples between 0.1 and 1 g/g was higher than 90%. The solid-phase adsorption technology resulted in a very effective methodology of clean-up in the case of the polar compound fluazifop, for which a second disposable column with a cyanopropyl phase has been used, and was fairly satisfactory for fluazifop-butyl. The detection limits were less than 0.04 g/g and 0.10 g/g, respectively for fluazifop and fluazifop-butyl.     

Simultaneous Determination of Fourteen Catecholamines and Their Metabolites by High Performance Liquid Chromatography with Electrochemical Detection

Abstract

A simple method has been developed for simultaneous determination of 14 catecholamines and their metabolites in cerebrospinal fluid and brain tissue by reversed-phase, ion-pair high-performance liquid chromatography with electrochemical detection. The time required for complete separation and analysis of all compounds was less than 35 min. Quantitation was based on the use of an internal standard isoproterenol. The mobile phase consisted of a 91:9 (v/v) mixture of 0.1 M formic acid and acetonitrile containing sodium-1-octane sulfonic acid. Using this method, analysis of neurotransmitters in brain tissue can be accomplished without a clean-up procedure.     

Improvement of selectivity and sensitivity by column switching in the determination of glycyrrhizin and glycyrrhetic acid in human plasma by high-performance liquid chromatography

A sensitive method is described for the simultaneous determination of glycyrrhizin and glycyrrhetic acid in human plasma. Quantitation is by high-performance liquid chromatography using ion-pair chromatography on a reversed-phase column. Variable-wavelength ultraviolet detection is employed. For the extraction of both compounds from plasma, a new solid-phase ion-pair extraction procedure using octadecylsilane columns was developed. Because of the strong forces involved in the protein binding of glycyrrhizin, quantitative extraction of this compound from plasma was possible only after an alkaline pH shift. A considerable improvement in selectivity and sensitivity was obtained by automated column switching involving on-line preseparation of the solid-phase extract on a short precolumn and chromatographic analysis of a heart-cut from the precolumn eluate. The limit of detection of both glycyrrhizin and glycyrrhetic acid was 0.1 mg/l in 0.5 ml of plasma. From a preliminary study in human volunteers, it was concluded that glycyrrhetic acid rather than glycyrrhizin is preferred in a study in human volunteers to assess the zero effect level of glycyrrhizin.     

LC-PDA and LC-ESI-MS separation and determination of process-related substances arising from stilbene-type fluorescent whitening agents. Application to monitoring of their photodegradation products in industrial effluents and aqueous environmental systems

A simple and rapid gradient elution high-performance liquid chromatographic method using photodiode array and electrospray ionization mass spectrometric detectors was developed for separation and determination of the process-related substances and photodegradation products of stilbenesulfonic acids, viz. 4,4'-dinitrostilbene-2,2'-disulfonic acid (DNSDA), 4-amino-4'-nitrostilbene-2,2'-disulfonic acid (ANSDA), and 4,4'-diaminostilbene-2,2'-disulfonic acid (DASDA) in industrial waste waters. Gradient elution was carried out using ammonium acetate and acetonitrile as mobile phase and an Inertsil-ODS 3V column for separation. The negative-ion electrospray ionization mass spectra containing [M-H]- ions of sulfonic acids allowed molecular mass determination of unknowns and the structures were proposed on the basis of the fragment ions in the MS/MS spectra.     

Determination of perfluoroalkyl carboxylic, sulfonic, and phosphonic acids in food

A sensitive and accurate method was developed and validated for simultaneous analysis of perfluoroalkyl carboxylic acids, sulfonic acids, and phosphonic acids (PFPAs) at low picograms per gram concentrations in a variety of food matrices. The method employed extraction with acetonitrile/water and cleanup on a mixed-mode co-polymeric sorbent (C8?+?quaternary amine) using solid-phase extraction. High-performance liquid chromatographic separation was achieved on a C18 column using a mobile phase gradient containing 5?mM 1-methyl piperidine for optimal chromatographic resolution of PFPAs. A quadrupole time-of-flight high-resolution mass spectrometer operating in negative ion mode was used as detector. Method detection limits were in the range of 0.002 to 0.02?ng?g^?1 for all analytes. Sample preparation (extraction and cleanup) recoveries at a spiking level of 0.1?ng?g^?1 to a baby food composite were in the range of 59 to 98?%. A strong matrix effect was observed in the analysis of PFPAs in food extracts, which was tentatively assigned to sorption of PFPAs to the injection vial in the solvent-based calibration standard. The method was successfully applied to a range of different food matrices including duplicate diet samples, vegetables, meat, and fish samples.

Determination of Pyridostigmine in Human Plasma by High-Performance Liquid Chromatography

Abstract

An easy to perform, specific, reproducible and sensitive high performance liquid chromatographic (HPLC) method to measure pyridostigmine concentration in human plasma was developed and validated. Sample clean-up consists of ion-pair extraction into dichloromethane in the presence of neostigmine as internal standard, followed by back extraction into an aqueous phase. Mean recovery of 110% (with a standard deviation of 10%) was determined for concentrations of 5 – 100 ng/ml. Chromatography on a 125·4 mm CN-propyl column using a mobile phase composed of 10% acetonitrile in 3.5×10^?4M NaH₂PO₄ and UV detection at 270 nm, yields clean chromatograms without any interferences from endogenous plasma components. Using 1 ml plasma samples the method has a limit of detection (LD) of 3 ng/ml, with %CV (precision) and bias (accuracy) ≥ 10% for concentrations in the range of 0–100 ng/ml. The method is being used in human pharmacokinetic studies of oral dosage forms of pyridostigmine.     

A rapid and environmental friendly determination of the dithiocarbamate metabolites ethylenethiourea and propylenethiourea in fruit and vegetables by ultra high performance liquid chromatography tandem mass spectrometry

Previous published methods for the analysis of ETU and PTU are time-consuming and furthermore use dichloromethane (DCM) for extraction or clean-up. This study details the development and validation of a rapid method that combines a simple extraction step with UHPLC-ESI(+)-MS/MS. This is the first application of UHPLC-MS/MS to analyse these compounds. Besides that, we replaced DCM with a more environmental-friendly solvent. The analytical performance was evaluated with the analysis of spiked celery samples at 50 μg kg(-1) (LOQ) and 300 μg kg(-1). The recoveries were between 65% and 90% for ETU and between 71% and 127% for PTU with RSDs in repeatability and reproducibility conditions below 10% for ETU. This method is rapid (a chromatographic run time of 2 min) and can easily be performed (no laborious clean-up). The presented method is environmental friendly with significant reduction in solvent consumption.     

Micelle formation for improvement of continuous subcritical water extraction of polycyclic aromatic hydrocarbons in soil prior to high-performance liquid chromatography-fluorescence detection

A new method for the extraction-individual separation-determination of polycyclic aromatic hydrocarbons (PAHs) in soil is reported. The method is based on the integration of three steps: continuous subcritical extraction, solid-phase clean-up/preconcentration, and HPLC separation with post-column fluorimetric determination. Sodium dodecyl sulfate (SDS) was added to the water for favouring the extractability of the low-polarity analytes. Soil samples spiked with the target PAHs were subjected to static-dynamic extraction with SDS-water at 50 bar, 150 degrees C, for 15 min of static extraction and 10 min dynamic extraction at a flow-rate of 3 ml/min. Recoveries from 73.6 to 110.4% were obtained in the presence of SDS versus 30 to 80% obtained with water as extractant. The calibration graphs provided by HPLC-fluorimetric detection were run between 0.031 and 0.375 microg/ml for each analyte with regression coefficients between 0.917 and 0.999 and precision, expressed as RSD, between 1.2 and 11.5%. The method was applied to a certified reference material [CRM 524, BCR (Community Bureau of Reference), industrial soil/organic] for validation and the results obtained were in agreement with the certified values.     

Method for the analysis in maize of the Fusarium mycotoxin moniliformin employing ion-pairing extraction and high-performance liquid chromatography

The Fusarium mycotoxin moniliformin (hydroxycyclobutenedione) has been determined in maize using a novel method with a recovery of 70-80% at 400-1600 micrograms/kg and 60% at the detection limit of 100 micrograms/kg. The method requires extraction of the toxin into aqueous tetra-n-butylammonium hydroxide and removal of cations from this solution by ion-exchange chromatography. Following clean-up by partitioning against dichloromethane, further quaternary ammonium reagent was added to the aqueous phase which was absorbed onto a hydrophilic matrix and the tetra-n-butylammonium moniliformate ion pair extracted into dichloromethane. After evaporation of the organic eluent, the residue was dissolved in aqueous sodium chloride and moniliformin quantitated by ion-pair high-performance liquid chromatography with UV detection. A batch of five samples may be analysed in 5-6 h including the chromatographic determination.     

Simultaneous determination of chromium (III) and chromium (VI) by reversed-phase ion-pair HPLC with chromium-specific detection

A method for the simultaneous determination of Cr(III) and Cr(VI) with reversed-phase ion-pair HPLC employing chromium-specific detection by flame atomic absorption spectrometry (FAAS) and inductively-coupled plasma mass spectrometry (ICP-MS) is presented. Experimental parameters of the chromatographic separation, such as concentration of the ion-pairing reagent, pH and polarity of the mobile phase have been optimized for two different ion-pairing reagents, tetrabutylammonium phosphate (TBA) and tetraethylammonium nitrate (TEA). Best chromatographic conditions have been obtained with a polymer-based reversed-phase column (Hamilton PRP1) and mobile phases containing either TBA (1 mmol/l) in methanol-water (60:40, v/v) or TEA (2 mmol/l) in water at a pH between 3 and 4. With FAAS the detection limits (3) have been found to be 24 g/l for Cr(III) and 40 g/l for Cr(VI). A detection limit of 0.3 g/l Cr(3) for both chromium species has been obtained when ICP-MS has been used for detection. The method has been applied to analyze tap- and groundwater and to investigate the behaviour of Cr(III) and Cr(VI) in spiked tap-water, as well as to analyze aqueous extracts of coal fly ash (NIST SRM 1633a) and of an ash from a wood treatment company.