Capillary electrophoretic determination of resveratrol in wines.

A rapid and sensitive capillary electrophoretic method for analysis of resveratrol in wine was established. The protocol consists of sample preparation using a C-18 solid-phase extraction cartridge. Baseline separation of trans- and cis-resveratrol from other compounds in wine was achieved in approximately 8 min using a micellar mode. The limits of detection for trans- and cis-resveratrol were 0.1 and 0.15 micromol/L, respectively. Recovery rates for trans-resveratrol using the protocol described ranged from 94.6 +/- 8.5 to 101.9 +/- 7.2%. These procedures were used to analyze the trans- and cis-resveratrol levels in 26 wines. It was found that the concentration of trans-resveratrol ranged from 0.987 to 25.4 micromol/L, whereas the concentration of cis-resveratrol was much lower.     

Piceid, the major resveratrol derivative in grape juices

The levels of trans-piceid, cis-piceid, trans-resveratrol, and cis-resveratrol have been measured in 36 grape juices using an HPLC system with spectral analysis of eluting peaks. The piceid (glucosides) were the major component in the grape juices. In red grape juices the average concentrations were 3.38 mg/L for trans-piceid, 0.79 mg/L for cis-piceid, 0.50 mg/L for trans-resveratrol, and 0.06 mg/L for cis-resveratrol. In white grape juices the levels were, on average, 0.18 mg/L for trans-piceid, 0.26 mg/L for cis-piceid, and 0.05 mg/L for trans-resveratrol, and cis-resveratrol was not detected in any sample. Levels of total resveratrol (trans- and cis-resveratrol and -piceid) found in red and in white grape juices are similar to those described in Spanish red and white wines. Due to their resveratrol content, as well as other phenolics, grape juices may have a beneficial health effect of interest to those who cannot drink wine.     

Development and validation of an analytical method for the determination of trans- and cis-resveratrol in wine: analysis of its contents in 186 Portuguese red wines

A simple procedure based on solid-phase extraction and high performance liquid chromatography coupled to diode array detector has been developed and validated for the qualitative and quantitative analysis of cis- and trans-resveratrol in wines. The method was linear from 0.025 (lower limit of quantitation, LLOQ) to 15 μg/mL for trans-resveratrol and from 0.023 (LLOQ) to 0.92 μg/mL for cis-resveratrol, with correlation coefficients higher than 0.99 for both isomers. Intra- and interday precision and accuracy were in conformity with the criteria normally accepted in method validation, that is, CVs inferior to 15% and mean relative errors within a ±14% interval. The extraction presented mean efficiencies close to 100% for both analytes. The validated methodology was applied to 186 Portuguese red wines from different regions, grape varieties and vintage. The results obtained showed that the content of trans-resveratrol in red wines ranged from 0.05 to 10.9 μg/mL, while the concentrations of cis-resveratrol ranged from 0.04 to 8.71 μg/mL.     

Determination of stilbenes (delta-viniferin, trans-astringin, trans-piceid, cis- and trans-resveratrol, epsilon-viniferin) in Brazilian wines

Phenolics from grapes and wines can play a role against oxidation and development of atherosclerosis. Stilbenes have been shown to protect lipoproteins from oxidative damage and to have cancer chemopreventive activity. We describe a method for the direct determination of stilbenes in several red wines using high-performance liquid chromatography with UV detection. In a survey of 12 commercial wines from the south of Brazil (Rio Grande del Sul), levels of delta-viniferin are reported for the first time in different varieties of red wines. Brazilian red wine contains trans-astringin, trans-piceid, trans-resveratrol, cis-resveratrol (in high quantity: 5 times more than the trans form), epsilon-viniferin, and a compound isolated for the first time in wine, trans-delta-viniferin. Isolation and identification of delta-viniferin was achieved by NMR after extraction and fractionation of red wine phenolics. delta-Viniferin contributes, as well as cis-resveratrol and trans-piceid, to a significant proportion of stilbenes in wine dietary intake, particularly with Merlot varieties containing an average level of 10 mg/L for delta-viniferin, 15 mg/L for cis-resveratrol, and 13 mg/L for trans-piceid. The total stilbene intake from wine origin was estimated for the Brazilian population as 5.3 mg/day per person (on the basis of a regular wine consumption of 160 mL/day). delta-Viniferin can contribute to around 20% of total stilbenes in wine (average of 6.4 mg/L in red Brazilian wines). It would be important in the future to investigate the origins of the differences in wine stilbene levels in relation to the vine varieties, and the bioavailability of the newly extracted stilbene delta-viniferin in plasma after consumption of different types of wines.     

Relationship among antioxidant activity, vasodilation capacity, and phenolic content of red wines

The relationship among antioxidant activity, based on the electron-spin resonance determination of the reduction of Fremy's radical, vasodilation activity, and phenolic content was investigated in 16 red wines. The wines were selected to provide a range of origins, grape varieties, and vinification methods. Sensitive and selective HPLC methods were used for the analysis of the major phenolics in red wine: free and conjugated myricetin, quercetin, kaempferol, and isorhamnetin; (+)-catechin, (-)-epicatechin, gallic acid, p-coumaric acid, caffeic acid, caftaric acid, trans-resveratrol, cis-resveratrol, and trans-resveratrol glucoside. Total anthocyanins were measured using a colorimetric assay. The total phenolic content of the wines was determined according to the Folin-Ciocalteu colorimetric assay and also by the cumulative measurements obtained by HPLC. The 16 wines exhibited a wide range in the values of all parameters investigated. However, the total phenol contents, measured both by HPLC and colorimetrically, correlated very strongly with the antioxidant activity and vasodilation activity. In addition, the antioxidant activity was associated with gallic acid, total resveratrol, and total catechin. In contrast, only the total anthocyanins were correlated with vasodilation activity. The results demonstrate that the different phenolic profiles of wines can produce varying antioxidant and vasodilatant activities, which opens up the possibility that some red wines may provide enhanced health benefits for the consumer.     

Analysis of coffee for the presence of acrylamide by LC-MS/MS

A variety of popular instant, ground, and brewed coffees were analyzed using a modified liquid chromatography-tandem mass spectrometry (LC-MS/MS) method specifically developed for the determination of acrylamide in foods. Coffee test portions were spiked with 13C3-labeled acrylamide as an internal standard prior to their extraction and cleanup. Ground coffees (1 g) and instant coffees (0.5 g) were extracted by shaking with 9 mL of water for 20 min. Brewed coffee test portions (9 mL) were taken through the cleanup procedure without further dilution with extraction solvent. Coffee test portions were cleaned up by passing 1.5 mL first through an Oasis HLB (hydrophilic/lipophilic copolymer sorbent) solid phase extraction (SPE) cartridge and then a Bond Elut-Accucat (cation and anion exchange sorbent) SPE cartridge. The cleaned up extracts were analyzed by positive ion electrospray LC-MS/MS. The MS/MS data was used to detect, confirm, and quantitate acrylamide. The limit of quantitation of the method was 10 ng/g for ground and instant coffees and 1.0 ng/mL for brewed coffee. The levels of acrylamide ranged from 45 to 374 ng/g in unbrewed coffee grounds, from 172 to 539 ng/g in instant coffee crystals, and from 6 to 16 ng/mL in brewed coffee.     

Determination of piceid and resveratrol in Spanish wines deriving from Monastrell (Vitis vinifera L.) grape variety

The presence of stilbenes in wine is becoming an important issue due to their claimed relation to a low incidence in coronary diseases and their increasing implication as cancer chemopreventive and neuroprotective agents. Total resveratrol content, quantified as glucoside and aglycone forms of resveratrol, has been determined in a survey of 45 Monastrell monovarietal Spanish red wine types (around 135 wine samples), belonging to Alicante and Bullas appellations. The average between ratio glucoside/aglycone forms of resveratrol in these wines was considerably high, ranging from 82 to 91% of resveratrol in its glycosidic form. This characteristic was observed in a high percentage of the studied wines, which were made under different winemaking procedures, and from different vintages (1995-2002). In addition, wines made using macerative fermentations with double amount of solid parts ("doble pasta") reached the highest levels of total stilbene content expressed as resveratrol equivalent, i.e., 30 mg/L (average of 18.8 mg/L). It can be concluded that high resveratrol glucoside concentration and low free isomer content can be considered characteristics of the Monastrell variety, as it happens to red wines deriving from other varieties grown at warm climates. This fact, also observed for other French and Portuguese red varieties, might play an important role in food habits involving these types of wines.     

Method for the gas chromatographic assay with mass selective detection of trichloro compounds in corks and wines applied to elucidate the potential cause of cork taint

To investigate the role of trichloro compounds as a potential cause of "cork taint" in wine, an assay for trichloroanisole (TCA) and trichlorophenol (TCP) in corks and wine was developed utilizing solid phase extraction on a C(18) cartridge followed by gas chromatography with mass selective detection. Recovery and imprecision for TCA were 86-102 and 1.6-5.8%, respectively, and for TCP 82-103% and 1.7-3.9%, respectively. Limits of detection and quantitation were 0.1 and 2 ng/L, respectively, for TCA, and 0.7 and 4 ng/L, respectively for TCP. A survey of 2400 commercial wines revealed a higher incidence of cork taint in white wine than in red and in wines utilizing composite cork closures; wines from central Europe and Spain had higher overall rates of contamination and those from Canada and Italy the lowest. Significant but modest associations were found between the TCA and TCP contents of the wines and corks, but many wines exhibiting cork taint had low or undetectable concentrations of TCA. Over a 12-month period, experimentally bottled wines exhibited a slow increase in TCA and TCP content while cork closures manifested a decrease; most bottles showing cork taint contained low levels of TCA, and TCP concentrations were well below the sensory threshold. Neither compound was cytotoxic to human cell lines in culture up to final concentrations of 500 ng/mL. It was concluded that these two trichloro compounds are, at most, minor components of cork taint in commercial wines.     

Analysis of ethyl carbamate in wines using solid-phase extraction and multidimensional gas chromatography/mass spectrometry

The method describes a rapid and accurate procedure for the analysis of ethyl carbamate in wines. The separation of the ethyl carbamate (EC), the target analyte, from alcohol and the sample matrix is a challenge to many analytical chemists. After alcohol removal from the sample, EC was extracted and concentrated by solid-phase extraction. For analysis of EC, large-volume injection on a programmable temperature vaporization (PTV) inlet was used followed by multidimensional gas chromatography/mass spectrometry (MDGC/MS) using electron-impact ionization (EI). For quantitation, the ratio of ions produced during EI at m/z 62 (EC) and 64 (isotopically labeled EC) was monitored. The use of solid-phase extraction and MDGC/MS removes the majority of the matrix interference encountered in other methods. A linear dynamic range was established from 0.387 to 1160 ng/mL, with a limit of detection at 0.1 ng/mL and limit of quantitation at 1 ng/mL.     

Occurrence of resveratrol in edible peanuts

Resveratrol has been associated with reduced cardiovascular disease and reduced cancer risk. This phytoalexin has been reported in a number of plant species, including grapes, and may be one of the compounds responsible for the health benefits of red wine. Analytical methods for measuring resveratrol in wine and peanuts were adapted to isolate, identify, and quantify resveratrol in several cultivars of peanuts. Aqueous ethanol (80% v/v) extracts from peanuts without seed coats were purified over alumina/silica gel columns and analyzed by reversed phase HPLC using a C-18 column. Peanuts from each market type, Virginia, runner, and Spanish, produced in four different locations contained from 0.03 to 0.14 microg of resveratrol/g. Seed coats from runner and Virginia types contained approximately 0.65 microg/g of seed coat, which is equivalent to <0.04 microg/seed. Quantitative analysis of 15 cultivars representing 3 peanut market types, which had been cold stored for up to 3 years, indicated a range of 0.02-1.79 microg/g of peanut compared to 0.6-8.0 microg/mL in red wines.     

Development of multiclass methods for drug residues in eggs: hydrophilic solid-phase extraction cleanup and liquid chromatography/tandem mass spectrometry analysis of tetracycline, fluoroquinolone, sulfonamide, and beta-lactam residues

A method was developed for detection of a variety of polar drug residues in eggs via liquid chromatography/tandem mass spectrometry (LC/MS/MS) with electrospray ionization (ESI). A total of twenty-nine target analytes from four drug classes-sulfonamides, tetracyclines, fluoroquinolones, and beta-lactams-were extracted from eggs using a hydrophilic-lipophilic balance polymer solid-phase extraction (SPE) cartridge. The extraction technique was developed for use at a target concentration of 100 ng/mL (ppb), and it was applied to eggs containing incurred residues from dosed laying hens. The ESI source was tuned using a single, generic set of tuning parameters, and analytes were separated with a phenyl-bonded silica cartridge column using an LC gradient. In a related study, residues of beta-lactam drugs were not found by LC/MS/MS in eggs from hens dosed orally with beta-lactam drugs. LC/MS/MS performance was evaluated on two generations of ion trap mass spectrometers, and key operational parameters were identified for each instrument. The ion trap acquisition methods could be set up for screening (a single product ion) or confirmation (multiple product ions). The lower limit of detection for screening purposes was 10-50 ppb (sulfonamides), 10-20 ppb (fluoroquinolones), and 10-50 ppb (tetracyclines), depending on the drug, instrument, and acquisition method. Development of this method demonstrates the feasibility of generic SPE, LC, and MS conditions for multiclass LC/MS residue screening.     

Automated on-line solid-phase extraction-liquid chromatography-electrospray tandem mass spectrometry method for the determination of ochratoxin A in wine and beer

An automated on-line solid-phase extraction-liquid chromatography-electrospray tandem mass spectrometry (SPE-LC-ESI-MS/MS) method was developed for the determination of ochratoxin A (OTA) in alcoholic beverages. Mean recoveries for wine and beer were, respectively, 75 and 82%. Detection was achieved in negative ionization with a Q TRAP mass spectrometer operating in multiple-reaction monitoring (MRM) mode or enhanced product ion (EPI) mode, using the third quadrupole as linear ion trap. The MRM mode turned out to be more sensitive; the method allowed accurate determination of OTA in the range of 0.01-25 ng mL(-1) using external calibration. Within-day and between-day relative standard deviation percentages were <6.2 and <9.1%, respectively. In EPI mode, fragmentation spectra at the limit of quantification (0.03 ng mL(-1)) and good linearity could be obtained. Application of the method (MRM mode) to the analysis of several wine and beer samples purchased in local stores revealed OTA levels in the ranges of 0.03-1.44 ng mL(-1) for wines and 0.02-0.14 ng mL(-1) for beers.     

Phenolic and volatile composition of wines made from Vitis vinifera cv. Muscat lefko grapes from the island of Samos

A study of the phenolic and volatile composition of wines produced from the white cultivar Muscat lefko from the island of Samos was conducted. Dry, fortified, naturally sweet wines and mistelles (aged and nonaged) have been studied. The phenolic components (flavan-3-ols, hydroxycinnamates, and flavonols) were measured by high-performance liquid chromatography after solid phase extraction (SPE). The terpenes (free and glycosidically linked) were determined by the use of gas chromatography-mass spectrometry (GC-MS) after SPE. The fermentation aroma components were analyzed by GC-MS after liquid-liquid extraction. It was found that the dry wines contained lower amounts of most of the phenolics and higher quantities of terpenes and fermentation aroma compounds than the sweet wines. The aged mistelle wines contained lower levels of coutaric and caftaric acids, higher concentrations of the free acids, and markedly fewer free and bound terpenes and fermentation aroma components compared to the other sweet wines. The naturally sweet wine contained relatively increased amounts of phenolics, 2,3-butanediol, and glycosidically linked terpenes.     

Determination of the phytoalexin resveratrol (3,5,4'-trihydroxystilbene) in peanuts and pistachios by high-performance liquid chromatographic diode array (HPLC-DAD) and gas chromatography-mass spectrometry (GC-MS)

The phytoalexin resveratrol (3,5,4'-trihydroxystilbene) in edible peanut (Arachis hypogaea L.) and pistachio (Pistacia vera L.) varieties grown in Turkey was analyzed by high-performance liquid chromatographic diode array and gas chromatography-mass spectrometric detection. trans-Resveratrol in six peanut varieties, five pistachio varieties, and four market samples ranged between 0.03 and 1.92 microg/g. The Cerezlik 5025 peanut (1.92 +/- 0.01 microg/g) and Ohadi pistachio genotype (1.67 +/- 0.01 microg/g) had significantly higher trans-resveratrol contents. Peanuts contained 0.03-1.92 microg/g (av = 0.84 microg/g) of trans-resveratrol, whereas pistachio contained 0.09-1.67 microg/g (av = 1.15 microg/g). With exposure to UV light for 1 min, trans-resveratrol concentrations of samples ranged from 0.02 to 1.47 microg/g and those of cis-resveratrol from 0.008 to 0.32 microg/g. The occurrence of resveratrol in peanut and pistachio was confirmed by total ion chromatograms (TIC) of bis[trimethylsilyl]trifluoroacetamide derivatives of resveratrol isomers and comparison of the mass spectral fragmentation data with those of a resveratrol standard. Formation of the cis-isomer in pistachios was higher than in peanuts.     

Ochratoxin a contamination in italian wine samples and evaluation of the exposure in the italian population

The scope of this study was to evaluate the exposure of the Italian population to ochratoxin A (OTA) attributable to wine consumption. With this aim 1166 wine samples (773 red wines, 290 white, 75 rose, and 28 dessert wines), collected in 19 different Italian regions and mostly produced between 1988 and 2004, were analyzed for OTA content. The obtained results are reported by year of harvest, geographical area of production, and type of wine. Red wine showed the highest maximum level of contamination (7.50 ng/mL), even though rose wines were characterized by a higher mean value (0.01 ng/mL). A gradually increasing mean concentration was also observed from the north (0.05 ng/mL) to south of Italy (0.54 ng/mL). Exposure calculations, performed using two different consumption databases, indicate a daily intake for consumer only of 0.59 up to 1.24 ng/(kg of b.w.)/day and of 0.33 up to 0.90 ng/(kg of b.w.)/day for the total population. Even in the worst case, corresponding to the calculation of the intake for consumers only in southern Italy and Islands and considering the mean consumption data increased by 1 standard deviation, a quite low exposure (1.68 ng/(kg of b.w.)/day, accounting for 9.8% of TDI) was obtained. Considering the overall OTA dietary exposure, obtained exposure rates indicate that wine did not pose a risk to the Italian population health.     

Resveratrol in raw and baked blueberries and bilberries

Resveratrol in the fruits of bilberry (Vaccinium myrtillus L.), the lowbush "wild" blueberry (Vaccinium angustifolium Aiton), the rabbiteye blueberry (Vaccinium ashei Reade), and the highbush blueberry (Vaccinium corymbosum L.) were measured using a new assay based on high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS). The LC-MS/MS assay provided lower limits of detection than previous methods for resveratrol measurement, 90 fmol of trans-resveratrol injected on-column, and a linear standard curve spanning >3 orders of magnitude. The recoveries of resveratrol from blueberries spiked with 1.8, 3.6, or 36 ng/g were 91.5 +/- 4.5, 95.6 +/- 6.5, and 88.0 +/- 3.6%, respectively. trans-Resveratrol but not cis-resveratrol was detected in both blueberry and bilberry samples. The highest levels of trans-resvertatrol in these specimens were 140.0 +/- 29.9 pmol/g in highbush blueberries from Michigan and 71.0 +/- 15.0 pmol/g in bilberries from Poland. However, considerable regional variation was observed; highbush blueberries from British Columbia contained no detectable resveratrol. Because blueberries and bilberries are often consumed after cooking, the effect of baking on resveratrol content was investigated. After 18 min of heating at 190 degrees C, between 17 and 46% of the resveratrol had degraded in the various Vaccinium species. Therefore, the resveratrol content of baked or heat-processed blueberries or bilberries should be expected to be lower than in the raw fruit. Although blueberries and bilberries were found to contain resveratrol, the level of this chemoprotective compound in these fruits was <10% that reported for grapes. Furthermore, cooking or heat processing of these berries will contribute to the degradation of resveratrol.     

Identification and quantification of stilbenes in fruits of transgenic tomato plants (Lycopersicon esculentum Mill.) by reversed phase HPLC with photodiode array and mass spectrometry detection

Reversed-phase high-performance liquid chromatography (RP-HPLC) with photodiode array (PDA) and mass spectrometry (MS) detection was employed to study the accumulation of stilbenes and other naturally occurring polyphenol intermediates of flavonoid pathway in tomato fruits of plants genetically modified to synthesize resveratrol. The transgenic tomato fruits were obtained by overexpression of a grapevine gene encoding the enzyme stilbene synthase in tomato plants (Lycopersicon esculentum Mill.). Stilbenes and flavonoids, either glycosylated or free, were simultaneosly identified by electrospray interface (ESI)-MS in negative ionization mode and were quantified by PDA detection at the wavelength corresponding to their maximum absorbance. The two detectors were coupled online with an HPLC system utilizing a narrow-bore C18 reversed-phase column, which was eluted by a multistep gradient of increasing concentration of acetonitrile in water containing 0.5% (v/v) formic acid. The results of these analysis revealed that the genetic modification of the tomato plants originated different levels of accumulation of four stilbenes (i.e., trans- and cis-piceid and trans- and cis-resveratrol) in their fruit depending on the stages of ripening. Either at immature or at mature stages of ripening the stilbenes were preferentially accumulated in the fruit peel as the glycosylated form. The highest amount of trans-piceid and trans-resveratrol were found in the peel of fruits harvested at mature stage of ripening. The variations in the levels of rutin, naringenin, and chlorogenic acid found in the samples extracted from the fruits of transgenic tomato plants, in comparison to that determined in the control lines, seemed to be related to the genetic transformation, whose effect on the flavonoid biosynthetic pathway needs to be elucidated by additional studies.     

Identification and quantification of geosmin, an earthy odorant contaminating wines

Musty, earthy odors are highly detrimental to the aromatic quality of wines. A characteristic aroma of freshly tilled earth, damp cellar was studied in some red and white wines of different origins. The extraction and purification of the wines marked by this odor have shown after analysis by gas chromatography-olfactometry a unique strong odorous zone having the same odor as the one perceived at tasting. The compound responsible for this odorous zone was identified by gas chromatography-mass spectrometry as geosmin (trans-1,10-dimethyl-trans-9-decalol), which possesses a distinctive earthy odor. Geosmin may be present in wines at levels higher than the racemic geosmin olfactory perception threshold, thus suggesting its contribution to their off-aroma. Moreover, the presence of this compound in juice taken from freshly crushed grapes suggests that microorganisms that develop on the grapes may contribute to the presence of this compound in wines.     

Stilbenes and tyrosol as target compounds in the assessment of antioxidant and hypolipidemic activity of Vitis vinifera red wines from southern Brazil

The contents of stilbene monomers, cis-resveratrol, trans-resveratrol, cis-piceid, trans-piceid, and tyrosol, were quantified in Vitis vinifera red wines, cvs. Cabernet Franc, Merlot, Sangiovese, and Syrah, 2006 and 2007 vintages, from the Sa?o Joaquim region, a new grape-growing region at southern Brazil. Moreover, the effect of chronic consumption of these wines on the antioxidant and hypolipidemic activities was monitored in C57BL6 LDL receptor knockout mice and treated with a hypercholesterolemic diet. Red wines from this region had substantial levels of resveratrols (the predominant forms were glycoside and trans) and tyrosol. Biomonitoring of antioxidant and hypolipidemic activities in vivo revealed that consumption of these wines increased the antioxidant capacity and reduced the hypercholesterolemia and hypertriglyceridemia promoted by the hypercholesterolemic diet. Significant correlations were found between the increase of antioxidant capacity markers, the decrease of lipid levels promoted by wine consumption, and the contents of stilbenes and tyrosol, supporting the important biological activity of these compounds.     

Simultaneous determination of ethidimuron, methabenzthiazuron, and their two major degradation products in soil

An analytical method has been developed for the quantification of two herbicides (ethidimuron and methabenzthiazuron) and their two main soil derivatives. This method involves fluidized-bed extraction (FBE) prior to cleanup and analysis by reverse-phase liquid chromatography with UV detection at 282 nm. FBE conditions were established to provide efficient extraction without degradation of the four analytes. (14)C-labeled compounds were used for the optimization of extraction and purification steps and for the determination of related efficiencies. Extraction was optimal using a fexIKA extractor operating at 110 degrees C for three cycles (total time = 95 min) with 75 g of soil and 150 mL of a 60:40 v/v acetone/water mixture. Extracts were further purified on a 500 mg silica SPE cartridge. Separation was performed on a C18 Purosphere column (250 mm x 4 mm i.d.), at 0.8 mL min(-1) and 30 degrees C with an elution gradient made up of phosphoric acid aqueous solution (pH 2.2) and acetonitrile. Calibration curves were found to be linear in the 0.5-50 mg L(-1) concentration range. Besides freshly spiked soil samples, method validation included the analysis of samples with aged residues. Recovery values, determined from spiked samples, were close to 100%. Limits of detection ranged between 2 and 3 microg kg(-1) of dry soil and limits of quantification between 8 and 10 microg kg(-1) of dry soil. An attempt to improve these performances by using fluorescence detection following postcolumn derivatization by orthophthalaldehyde-mercaptoethanol reagent was unsuccessful.