Rapid method for determination of phenylurea herbicides in milk

Summary A method has been developed for the determination of thirteen phenylurea herbicide residues in milk. It involves one-step solvent extraction of the milk with methanol by ultrasonication. The extract is cleaned up on an Amberchrom resin cartridge. Reversed-phase, gradient elution, high-performance liquid chromatography with UV detection at 242 nm is used to analyse the residues. The recovery of thirteen phenylurea pesticides is quantitative, ranging from 71.4% to 97.9% for the individual herbicides investigated at concentrations around 0.05 mg kg^–1 and from 65.1% to 95.6% around 0.005 mg kg^–1. The method is not associated with any of the emulsion problems common to conventional solvent extraction, which considerably reduce the sample clean-up process compared with existing methods.     

Application of an innovative matrix solid-phase dispersion–solid-phase extraction–liquid chromatography–tandem mass spectrometry analytical methodology to the study of the metabolism of the estrogenic mycotoxin zearalenone in rainbow trout liver and muscular tissue

The metabolic fate of the estrogenic mycotoxin zearalenone in rainbow trout is presently unknown. In this study, the tissue concentration of zearalenone and its principal metabolites (α-zearalenol and β-zearalenol) was determined. A known amount of zearalenone was administered as a single bolus to ten fish, and the biological tissue concentration was determined at various times following administration. The analytes were extracted from liver and muscular tissue using an on-line matrix solid-phase dispersion–solid-phase extraction sample preparation protocol, and their concentration determined by HPLC–Turboionspray–tandem mass spectrometry. The results showed that zearalenone is mainly metabolized into α-zearalenol in both liver and muscular tissues. The maximum concentrations of each analyte found in liver were 76.1, 211.2 and 63.7?ng/g respectively for zearalenone, α-zearalenol and β-zearalenol, while in muscular tissue they were 10.7, 8.2 and 6.5?ng/g. These values were reached after 2?h in liver tissue and 12?h in muscular tissue. Moreover the data obtained showed that the elimination rate in liver is quite fast since 48?h after the exposure less than 7% of the maximum concentration found is still present. In muscular tissue, however, about one-third of the maximum concentration found is still present after 48?h.     

Determination of maize and grain herbicides and their transformation products in soil by use of soil column extraction then liquid chromatography with tandem mass spectrometry

Summary A multiresidue method has been developed for identification and quantitation of the herbicides most commonly used in cultivation of maize and grain, and of their transformation products, in soil samples. The analytes were isolated by soil column extraction (SCE) and the extracts were purified by use of a Carbograph-1 cartridge. Analysis was performed by liquid chromatography coupled with tandem mass spectrometric detection (LC-TISP-MS-MS) in negative-ion mode. To optimize the extraction conditions affecting the performance of SCE, e.g. the extracting solvent used, temperature, extracting volume, and solvent flow rate, were studied. To evaluate the matrix effect in SCE, recovery experiments were performed on soil samples, with different physical and chemical characteristics, fortified with the 100 ng g⁻¹ of the target compounds. Recovery data were satisfactory and the method detection limits were between 3 and 100 ng g⁻¹, depending on the compound.     

Chromatographic purification and HPLC assay with electrochemical detection of diethylstilboestrol residues in meat

Summary A rapid, reliable and sensitive procedure is reported for the detection of residues of total diethylstilboestrol (DES) (free and conjugated) in bovine muscle samples. Prior to analysis, the DES in the sample was enzymatically deconjugated. Subsequently the sample was subjected to a clean-up procedure using a two-trap system composed of a first cartridge of non-specific adsorbing material (C 18) and of a second cartridge containing a sulphonic acid type silica-based anion exchanger. The determination was performed by HPLC with electrochemical detection. Analytical variables such as efficiency of hydrolysis, enzyme purification, hydrodynamic voltammetric studies and variation in the DES trans/cis ratio were investigated. Mean recovery of total DES was 87.3%. The precision of the procedure was 7.2% for DES levels of 0.5 ng/g and 2.1% for levels of 5 ng/g.     

An improved method for the determination of free sphingosine in serum

Summary A rapid, sensitive and selective high-performance liquid chromatographic method has been developed for the determination of sphingosine in human serum. After precipitation with methanol, the samples were extracted using Carbopack B disposable columns; the sphingosine was eluted with 0.05 M hydrochloric acid in methanol-dichloromethane (20∶80, v/v) and the extract evaporated to dryness at 40°C. The sample residue was then reconstituted with methanol and reacted with o-phthaldialdehyde reagent to produce a fluorescent compound. Separation was performed using an LC-18 column with 0.05 M phosphate buffer (pH 7)-methanol-acetonitrile (15∶80∶5, v/v) as mobile phase. Fluorescence detection was performed with excitation and emission wavelengths of 340 and 455 nm, respectively. The serum extract was re-analyzed with a cyano LC column to minimize the possibility of false positive results. The possible interference of compounds having a structure similar to that of sphingosine was evaluated. The mean recovery of sphingosine was >94.5%. The limit of detection of the assay was 1 ng mL⁻¹. The between-run and within-run coefficients of variation for replicate analyses were <4.0% and <3.4%, respectively. The levels of free sphingosine in the serum of 40 normal subjects (20 male and 20 female) was investigated; the average level was 81.6±41.1 ng mL⁻¹ (mean ±S.D.) for males and 85.5±33.7 ng mL⁻¹ for females.     

Trace analysis of estrogenic chemicals in sewage effluent using liquid chromatography combined with tandem mass spectrometry

A rapid, selective, sensitive, and reproducible method for the analysis of environmental estrogens, either natural or synthetic, present in samples from sewage treatment works (STW) has been developed. Isolation of these compounds from STWs was performed by applying a simple extraction procedure using an ENVI-CARB cartridge (a graphitized non porous carbon black) as the solid-phase extraction (SPE) system. Liquid chromatography coupled with atmospheric pressure chemical ionization (APCI) and tandem mass spectrometry was used for detection. For the multiple reaction monitoring (MRM) mode, the [M + H](+) ion of estrone and the [M + H-H(2)O](+) ions of 17beta-estradiol, estriol and 17alpha-ethynylestradiol were selected as the precursors for collisionally induced dissociation (CID). The average recoveries from sewage final effluent samples ranged from 84 to 93% for low levels, and from 89 to 95% for high levels. The precision of the method ranged from 11 to 8% for low level and from 9 to 7% for high level samples. The lower level of quantitation for these estrogens in STW samples was determined at 0.5 ng/L for 17beta-estradiol and 17alpha-ethynylestradiol, and 1 ng/L for estrone and estriol, based on 1-L aliquots of sewage treatment works water, using the optimum tuning parameters for each individual selected precursor ion/product ion transition. Compared to a previous gas chromatography/mass spectrometry (GC/MS) method for the analysis of ten STW samples, this method was shown to provide higher sensitivity and lower time consumption.     

Evaluation of double solid-phase extraction system for determining triazine herbicides in milk

Summary High-performance liquid chromatography with UV detection was used to determine eight triazine herbicides in milk. Solid-phase extraction was performed using a double trap; first, a nonspecific adsorbent (Carbograph), and then a cation exchanger (SCX).Eluate from the SCX was evaporated to dryness under reduced pressure and redissolved in mobile phase. An aliquot was injected into the chromatography, which was operated isocratically in the reverse-phase mode with UV detection at 225 nm.Analytical recoveries for the eight triazines ranged from 73.0% to 92.4%. The limit of sensitivity of this method was about 0.09 ng mL^–1 of milk. The method was validated and evaluated by comparison with a method reported in literature.     

Development of an analytical system for the simultaneous determination of anabolic macrocyclic lactones in aquatic environmental samples

An analytical procedure that enables routine analysis for trace determination of six anabolic macrocyclic lactones (zearalenone, alpha- and beta-zearalenol, zearalanone, zeranol, and taleranol) in sewage treatment plant (STP) samples has been developed. The method uses solid-phase extraction, followed by high-performance liquid chromatography with on-line tandem mass spectrometry using atmospheric pressure chemical ionization (LC/APCI-MS/MS). The extraction of these compounds from filtered water samples was performed off-line with C(18) solid-phase cartridges. The detection was achieved by isocratic reversed-phase high-performance liquid chromatography coupled with an heated nebulizer (HN) APCI interface operating in negative ion mode. Mean recovery of the analytes in STP effluent samples generally exceeded 81%. This method was used to determine the occurrence of target analytes in the aquatic environment. In the selected STP effluent samples, zearalenone and alpha-zearalenol were detected in the ng/L range.