Pilar Viñas

Determination of phenols in wines by liquid chromatography with photodiode array and fluorescence detection

A reversed-phase LC method, optimised for the separation of trans- and cis-resveratrol, catechin, epicatechin, quercetin and rutin, is reported. Analyses were performed on a reversed-phase column by gradient elution. Detection was carried out by photodiode array, although the use of a fluorimetric detector considerably lowered the detection limits for catechin, epicatechin and both resveratrol isomers. Identification by the two different detection systems was based on retention characteristics, UV spectra and peak purity index were compared with commercial standards. The procedures were applied to the determination of the phenolic compounds in different types of wines and musts.

Dispersive liquid–liquid microextraction in food analysis. A critical review

An extensive critical evaluation of the application of dispersive liquid–liquid microextraction (DLLME) combined with chromatographic and atomic-spectroscopic methods for the determination of organic and inorganic compounds is presented. The review emphasizes the procedures used for the prior treatment of food samples, which are very different from the DLLME procedures generally proposed for water samples. The main contribution of this work in the field of DLLME reviews is its critical review of the abundant literature showing the increasing interest and practical advantages of using DLLME and closely related microextraction techniques for food analysis.

Reversed-phase liquid chromatography on an amide stationary phase for the determination of the B group vitamins in baby foods.

A liquid chromatographic method for the separation and determination of several water-soluble vitamins of the B group is proposed. The procedure is based on the use of a new amide-based stationary phase, which avoids the need of using the ion-pair technique, leading to narrower peaks and a simpler mobile phase. Analyses were performed by gradient elution with acetonitrile-phosphate buffer as mobile phase and using a photodiode array detector. Specificity was demonstrated by the retention characteristics, UV spectra and by comparing the peak purity index with commercial standards. Linearity, precision, recovery and sensitivity were satisfactory. The vitamins are extracted from the baby food using a single digestion with hydrochloric acid followed by enzymatic digestion with taka-diastase. The method was successfully applied to the determination of nine vitamins: thiamine, riboflavin, nicotinamide, nicotinic acid, pyridoxine, pyridoxal, folic acid, cyanocobalamine and inosine in different baby foods as infant formulas, cereals and fruit products. Validation was performed using two certified reference materials.

Method development and validation for strobilurin fungicides in baby foods by solid-phase microextraction gas chromatography-mass spectrometry.

The present study describes a new solvent-free method for the sensitive determination of seven strobilurin fungicides (azoxystrobin, metominostrobin, kresoxim-methyl, trifloxystrobin, picoxystrobin, dimoxystrobin and pyraclostrobin) in baby food samples. Direct immersion solid-phase microextraction (DI-SPME) coupled to gas chromatography with mass spectrometry in the selected ion monitoring mode, GC-MS (SIM), is used. All analyses were performed with 2g of sample mass, 14mL of sample extract volume and sample extract buffered at pH 5. Optimal extraction conditions were 60 degrees C for 40min under continuous stirring using a PDMS-DVB fiber. Desorption was carried out at 240 degrees C for 4min. The standard additions method is recommended and quantitation limits ranged from 0.01 to 0.4ngg(-1) at a signal to noise ratio of 10, depending on the compound. Recoveries obtained for spiked samples were satisfactory for all the compounds. The method was validated according to the Commission Decision 2002/657/EC. Different baby foods were analyzed by the proposed method and none of the samples contained residues above the detection limits.

Placental lead and outcome of pregnancy.

Prenatal exposure to lead produces toxic effects in the human fetus, including an increased risk of preterm delivery, low birth weight and impaired mental development. Since we believe that placental lead could be a good biomarker for fetal exposure, we assessed the relation of placental lead to intrauterine fetal growth and some adverse outcomes of pregnancy. Low concentrations of lead were found in the population studied (N, 89; mean, 113.4 ng/g dry tissue; S.D., 58.0). The cases were divided into two groups based on delivery outcome. In the group of placentas from premature rupture of membranes and preterm labor (gestational age <or=37 weeks), higher lead levels were recorded than in the placentas from term pregnancies. The proportion of abnormal pregnancy outcome in the group of placentas with lead concentrations above 120 ng/g was 40.6 versus 8.8% in placentas below this concentration. Higher placental lead levels, in general, were not related to smaller weight, head and abdominal circumference or shorter length at birth.

Determination of volatile nitrosamines in meat products by microwave-assisted extraction and dispersive liquid–liquid microextraction coupled to gas chromatography–mass spectrometry

Microwave-assisted extraction (MAE) and dispersive liquid-liquid microextraction (DLLME) coupled with gas chromatography-mass spectrometry (GC-MS) were evaluated for use in the extraction and preconcentration of volatile nitrosamines in meat products. Parameters affecting MAE, such as the extraction solvent used, and DLLME, including the nature and volume of the extracting and disperser solvents, extraction time, salt addition and centrifugation time, were optimized. In the MAE method, 0.25g of sample mass was extracted in 10mL NaOH (0.05M) in a closed-vessel system. For DLLME, 1.5mL of methanol (disperser solvent) containing 20μL of carbon tetrachloride (extraction solvent) was rapidly injected by syringe into 5mL of the sample extract solution (previously adjusted to pH 6), thereby forming a cloudy solution. Phase separation was performed by centrifugation, and a volume of 3μL of the sedimented phase was analyzed by GC-MS. The enrichment factors provided by DLLME varied from 220 to 342 for N-nitrosodiethylamine and N-nitrosopiperidine, respectively. The matrix effect was evaluated for different samples, and it was concluded that sample quantification can be carried out by aqueous calibration. Under the optimized conditions, detection limits ranged from 0.003 to 0.014ngmL(-1) for NPIP and NMEA, respectively (0.12-0.56ngg(-1) in the meat products).

Stir bar sorptive extraction coupled to gas chromatography-mass spectrometry for the determination of bisphenols in canned beverages and filling liquids of canned vegetables.

This paper describes a method for the simultaneous determination of bisphenol A (BPA), bisphenol F (BPF), bisphenol Z (BPZ) and biphenol (BP), using stir bar sorptive extraction (SBSE) in combination with thermal desorption-gas chromatography-mass spectrometry (TD-GC-MS). Several parameters affecting both extraction and thermal desorption of the SBSE stages were carefully optimized by multivariate designs. SBSE was performed with two derivatization procedures, in situ acetylation and in tube silylation, and the results were compared with those obtained when the analytes were not derivatized. The proposed method, determining the analytes as acyl derivatives, was applied to analyze commercially canned beverages, as well as the filling liquids of canned vegetables, providing detection limits of between 4.7 and 12.5 ng L⁻¹, depending on the compound. The intraday and interday precisions were lower than 6% in terms of relative standard deviation. Recovery studies at two concentration levels, 0.1 and 1 μg L⁻¹, were performed providing recoveries in the 86-122% range. The samples analyzed contained higher concentrations of BPA than of the other analytes.

Determination of alkylphenols and phthalate esters in vegetables and migration studies from their packages by means of stir bar sorptive extraction coupled to gas chromatography–mass spectrometry

This paper describes a method for the determination of three alkylphenols (APs), 4-tert-octylphenol (tOP), 4-n-octylphenol (OP) and 4-nonylphenol (NP), and six phthalate esters (PEs), dimethylphthalate (DMP), diethylphthalate (DEP), di-n-butylphthalate (DBP), n-butylbenzylphthalate (BBP), di-2-ethylhexylphthalate (DEHP) and di-n-octylphthalate (DOP), in vegetables using stir bar sorptive extraction (SBSE) in combination with thermal desorption-gas chromatography-mass spectrometry (TD-GC-MS). Ultrasonic radiation was used to extract the analytes from the solid food matrix, and the extract obtained was preconcentrated by SBSE. The different parameters affecting both stages were carefully optimized. The method was applied to analyze commercial vegetables, in the form of plastic packed salads and canned greens, as well as the corresponding filling liquids of the canned food. Quantification of the samples was carried out against aqueous standards using an internal standard (anthracene). The analysis of a 2 g vegetable sample provided detection limits between 12.7 and 105.8 pg g⁻¹ for OP and DEHP, respectively. Migration studies from the plastic packages of the vegetables samples analyzed were carried out. DEP, DBP and DEHP were found to have migrated from the bags to the simulant and the same compounds were quantified in lettuce, corn salad, arugula, parsley and chard, at concentration levels in the 8-51 ng g⁻¹ range. However, OP and NP were found in only two vegetable samples and one filling liquid, but neither was detected in any package. The proposed method provided recoveries of 83-118%.

Solid-phase microextraction on-fiber derivatization for the analysis of some polyphenols in wine and grapes using gas chromatography–mass spectrometry

The present study describes a new environmentally friendly sample pretreatment system based on solid-phase microextraction (SPME) for the sensitive determination of polyphenols. A derivatization process was necessary to convert the polar non-volatile compounds into volatile derivatives. Direct immersion (DI) SPME was used for the adsorption of polyphenols, and then the fiber was placed in the headspace of the derivatizing reagent, bis(trimethylsilyl)trifluoroacetamide (BSTFA). The separation was carried out by coupling gas chromatography with mass spectrometry in the selected ion monitoring mode, after silylation. Optimal extraction conditions were 25 degrees C for 10 min under continuous stirring using DI and a polyacrylate fiber. After extraction, the fiber was inserted into the headspace of BSTFA (10 microL) and the polyphenols were derivatized for 15 min at 50 degrees C. Desorption was carried out at 280 degrees C for 5 min. The method allowed the determination of both isomers cis- and trans-resveratrol, piceatannol, catechin and epicatechin in wine and grapes, and it was validated for linearity, detection and quantitation limits, selectivity, accuracy and precision. Detection limits ranged from 0.05 to 0.9 ng mL(-1) at a signal-to-noise ratio of 3, depending on the compound. Recoveries obtained for spiked samples were satisfactory for all compounds.

Evaluation of dispersive liquid–liquid microextraction for the simultaneous determination of chlorophenols and haloanisoles in wines and cork stoppers using gas chromatography–mass spectrometry

Dispersive liquid-liquid microextraction (DLLME) coupled with gas chromatography-mass spectrometry (GC-MS) was evaluated for the simultaneous determination of five chlorophenols and seven haloanisoles in wines and cork stoppers. Parameters, such as the nature and volume of the extracting and disperser solvents, extraction time, salt addition, centrifugation time and sample volume or mass, affecting the DLLME were carefully optimized to extract and preconcentrate chlorophenols, in the form of their acetylated derivatives, and haloanisoles. In this extraction method, 1mL of acetone (disperser solvent) containing 30μL of carbon tetrachloride (extraction solvent) was rapidly injected by a syringe into 5mL of sample solution containing 200μL of acetic anhydride (derivatizing reagent) and 0.5mL of phosphate buffer solution, thereby forming a cloudy solution. After extraction, phase separation was performed by centrifugation, and a volume of 4μL of the sedimented phase was analyzed by GC-MS. The wine samples were directly used for the DLLME extraction (red wines required a 1:1 dilution with water). For cork samples, the target analytes were first extracted with pentane, the solvent was evaporated and the residue reconstituted with acetone before DLLME. The use of an internal standard (2,4-dibromoanisole) notably improved the repeatability of the procedure. Under the optimized conditions, detection limits ranged from 0.004 to 0.108ngmL(-1) in wine samples (24-220pgg(-1) in corks), depending on the compound and the sample analyzed. The enrichment factors for haloanisoles were in the 380-700-fold range.