Variability in chemical composition of Vitis vinifera cv Mencía from different geographic areas and vintages in Ribeira Sacra (NW Spain).

A chemical study was conducted from 2009 to 2012 to examine spatial and seasonal variability of red Vitis vinifera Mencía located in different geographic areas (Amandi, Chantada, Quiroga-Bibei, Ribeiras do Sil and Ribeiras do Miño) from NW Spain. Mencía samples were analysed for phenolic, (flavan-3-ols, flavonols, anthocyanins, acids and resveratrol), nitrogen (TAC, TAN, YAN and TAS) and volatiles compounds (alcohols, C6 compounds, ethyl esters, terpenes, aldehydes, acids, lactones, volatile phenols and carbonyl compounds) by GC-MS and HPLC. Results showed that the composition of Mencía cultivar was more affected by the vintage than the geographic area. The amino acid composition was less affected by both geographic origin and vintage, showing more varietal stability. Application of Principal Component Analysis (PCA) to experimental data showed a good separation of Mencía grape according to geographical origin and vintages. PCA also showed high correlations between the ripening ratio and C6 compounds, resveratrol and carbonyl compounds.

Conversion of heart transplant patients from standard to sustained-release tacrolimus requires a dosage increase.

INTRODUCTION: It has been suggested that for adequate maintenance of tacrolimus levels, the total daily dosage should be increased when switching from the conventional twice-daily regimen tacrolimus (CT) to once-daily sustained-release tacrolimus (SR-T). OBJECTIVE: To evaluate the safety and efficacy of a 25% increase in daily dosage when switching heart transplant (HT) patients from CT to SR-T. METHODS: We switched 75 HT patients including 72% males and an overall mean age of 55.6 years from CT to SR-T using a 25% increase in daily dosage. We screened for adverse events by measurements of lipids, creatinine, glycemia, and tacrolimus in blood samples taken at 1, 3, 7, and 12 weeks after the conversion, as well as by repeated echocardiography and routine clinical examinations. RESULTS: Just two patients (2.7%) were returned to CT because of failure of SR-T to attain therapeutic levels. In the remainder of subjects, tacrolimus levels remained stable, with trough values of 8.7±3.2, 8.7±2.9, 8.3±2.6, and 7.5±2.0 mg/dL, respectively. Twenty-three patients (31%) required no dosage change in the first 3 months, but 44 (33%) required one or two changes. No departure from therapeutic levels was associated with rejection; there was no case of severe intercurrent infection. We did not observe significant changes in glycemia, creatinine, lipid profile, or blood pressure. CONCLUSIONS: Administration of SR-T at a dosage 25% higher than the daily dosage of CT was safe. It ensured adequate tacrolimus levels in one-third of patients. Nevertheless, strict analytical surveillance is necessary during the initial months to allow dosage adjustments and to detect the minority of patients for whom SR-T does not achieve therapeutic tacrolimus levels.

Matrix solid-phase dispersion and solid-phase microextraction applied to study the distribution of fenbutatin oxide in grapes and white wine.

The fate of the acaricide fenbutatin oxide (FBTO) during the elaboration of white wine is evaluated. Matrix solid-phase dispersion (MSPD) and solid-phase microextraction (SPME) were used as sample preparation techniques applied to the semi-solid and the liquid matrices involved in this research, respectively. Selective determination of FBTO was achieved by gas chromatography with atomic emission detection (GC-AED). GC coupled to mass spectrometry was also used to establish the identity of FBTO by-products detected in must and wine samples. MSPD extractions were accomplished using C18 as dispersant and co-sorbent. Sugars and other polar interferences were first removed with water and water/acetone mixtures, then FBTO was recovered with 8 mL of acetone. When used in combination with GC-AED, the MSPD method provided limits of quantification (LOQs) in the low nanogram per gram range, recoveries around 90% and relative standard deviations below 13% for extractions performed in different days. Performance of SPME for must and wine was mainly controlled by the extraction temperature, time and fibre coating. Under final conditions, FBTO was extracted in the headspace mode for 45 min at 100 degrees C, using a 100 microm poly(dimethylsiloxane)-coated fibre. The achieved LOQs remained around or below 0.1 ng mL(-1), depending on the type of sample, and the inter-day precision ranged from 10% to 13%. FBTO residues in grapes stayed mostly on the skin of the fruit. Although FBTO was not removed during must and white wine elaboration, it remained associated with suspended particles existing in must and lees, settled after must fermentation, with a negligible risk of being transferred to commercialised wine. On the other hand, two by-products of FBTO (bis and mono (2-methyl-2-phenylpropyl) tin) were identified, for first time, in must and final white wines obtained from FBTO treated grapes. Found values for the first species ranged from 0.03 to 0.9 ng mL(-1).

Pressurized liquid extraction followed by gas chromatography with atomic emission detection for the determination of fenbutatin oxide in soil samples.

A novel method for the determination of the miticide bis[tris(2-methyl-2-phenylpropyl)tin] oxide, also known as fenbutatin oxide (FBTO), in agricultural soils is presented. Pressurized liquid extraction (PLE) followed by analyte derivatization and extraction into isooctane was the used sample preparation approach. Selective determination was achieved by gas chromatography with atomic emission detection (GC-AED). Influence of different parameters on the performance of the extraction process is thoroughly discussed; moreover, some relevant aspects related to derivatization, determination and quantification steps are also presented. As regards PLE, the type of solvent and the temperature were the most relevant variables. Under optimized conditions, acetone, without any acidic modifier, was employed as extractant at 80 degrees C. Cells were pressurized at 1500 psi, and 2 static cycles of 1 min each were applied. Acetone extracts (ca. 25 mL) were concentrated to 1 mL, derivatized with sodium tetraethyl borate (NaBEt(4)) and the FBTO derivative, resulting from cleavage of the Sn-O-Sn bond followed by ethylation of the hydroxyl fragments, extracted into isooctane and determined by GC-AED. Under final working conditions, the proposed method provided recoveries from 76 to 99% for spiked soil samples, a limit of quantification of 2 ng g(-1) and an acceptable precision. Analysis of samples from vineyards sprayed with FBTO, confirmed the persistence of the miticide in soil for more than 1 year after being applied.

Simplified sample preparation method for triclosan and methyltriclosan determination in biota and foodstuff samples.

An improved method for the determination of triclosan (TCS) and methyltriclosan (MTCS) in fish and foodstuff samples is presented. Analytes were simultaneously extracted and purified using the matrix solid-phase dispersion (MSPD) technique, and then selectively determined by gas chromatography with tandem mass spectrometry (GC-MS/MS). Several combinations of dispersants, clean-up co-sorbents and extraction solvents were tested in order to obtain lipid-free extracts and quantitative recoveries for TCS and MTCS. Under optimised conditions, 0.5 g samples were dispersed using 1.5 g of neutral silica in a mortar with a pestle, and transferred to a polypropylene cartridge containing 3 g of silica impregnated with 10% of sulphuric acid (SiO2-H2SO4, 10%, w/w). Analytes were recovered with 10 mL of dichloromethane whereas lipids were oxidized in the layer of acidic silica. The extract was concentrated to dryness and re-constituted with 1 mL of ethyl acetate. Then, a fraction of 0.5 mL was mixed with 50 microL of N-methyl-N-(tert-butyldimethylsilyl)trifluoroacetamide (MTBSTFA) and injected in the GC-MS/MS system. The developed method provided absolute recoveries between 77 and 120% for different samples spiked at the low ng g(-1) level, quantification limits in the range of 1-2 ng g(-1) and a considerable simplicity in comparison with previously developed sample preparation approaches. Experiments carried out placing sliced food samples in direct contact with TCS-treated kitchenware surfaces showed the capability of the biocide to migrate into foodstuffs.

Trends and recent applications of matrix solid-phase dispersion.

Matrix solid-phase dispersion (MSPD) is a sample-preparation technique with increasing acceptance in trace analysis of organic compounds using chromatographic and electro-driven separation techniques. It has been applied to the extraction and fractionation of a large number of substances from solid, semi-solid, and liquid matrices. Low sample and solvents consumption, straightforward application, and reduced cost, and its ability to simultaneously perform extraction and clean-up in a single step, are some of its major advantages. This review attempts to provide an updated, concise and critical overview on the latest trends and applications of MSPD, placing emphasis on comparison of its performance with that of other techniques, besides focusing on practical features to take into account depending on the nature of the sample and the properties of the analytes. Achievements, advantages, and limitations are discussed. The paper also highlights future challenges to be faced.

Study of some UV filters stability in chlorinated water and identification of halogenated by-products by gas chromatography-mass spectrometry.

This work studies the stability of three UV filters: 2-ethylhexyl salicylate (ES), 2-ethylhexyl 4-(dimethylamino) benzoate (EHPABA) and 2-hydroxy-4-methoxybenzophenone (BP-3), in water samples containing low concentrations of free chlorine. Moreover, 2,4-dihydroxybenzophenone (2,4-DHBP), a metabolite of BP-3, was also included in some of the performed assays. Experiments were carried out considering free chlorine and analytes concentrations at the microg mL(-1) and ng mL(-1) level, respectively. Gas chromatography with mass spectrometry was used to follow the time course of target compounds and to identify their halogenated by-products. Concentration of water samples with solid-phase extraction cartridges and derivatization (silylation) of some species were also employed to improve their detectability. Under the experimental conditions explored in this work, ES showed an acceptable stability whereas the rest of species reacted with free chlorine at significant rates following pseudo-first-order kinetics. Their half-lives ranged from 0.4 to 25 min depending on the UV filter, chlorine concentration, water pH and presence of bromide traces. For EHPABA a relatively simple degradation pathway was established. It consisted of aromatic substitution of one atom of hydrogen per chlorine or bromide. The same reaction pattern was observed for BP-3 leading, in this case, to mono- and di-halogenated by-products. In addition, several halogenated forms of 3-methoxyphenol were identified as BP-3 cleavage by-products.

Pressurized liquid extraction with in-cell clean-up followed by gas chromatography-tandem mass spectrometry for the selective determination of parabens and triclosan in indoor dust.

A sample preparation method based on the use of pressurized liquid extraction is proposed for the determination of four alkyl parabens and triclosan in indoor dust. Extraction of analytes and removal of interfering species were achieved in the same step, by placing an appropriate sorbent in the extraction cell and by choosing a right combination of washing and elution solvents. Compounds, as silylated derivatives, were determined by gas chromatography in combination with tandem mass spectrometry (GC-MS/MS). Factors affecting the yield and selectivity of the sample preparation procedure were carefully evaluated. Under final conditions, dried samples (0.5 g of dust and 1g of sodium sulphate) were dispersed with 3g of Florisil and loaded into an 11 mL stainless-steel extraction cell containing 1g of the same material as clean-up sorbent. Non-polar species were removed with n-hexane under mild conditions (40 degrees C, 3.4 MPa) and then analytes were extracted with ethyl acetate. The best compromise extraction conditions were 103 degrees C, 13.8 MPa and 3 static extraction cycles of 1 min. The proposed method provided recoveries from 76 to 98%, relative standard deviations under 11% (operating under reproducibility conditions) and quantification limits from 1 to 4 ng/g. The analysis of dust samples from private houses and office buildings confirmed the ubiquitous presence of target bacteriocides in these environments.

Determination of parabens and triclosan in indoor dust using matrix solid-phase dispersion and gas chromatography with tandem mass spectrometry.

A simple sample preparation method for the determination of four parabens and triclosan in indoor dust is presented. Analytes were extracted from the sample and isolated from interfering species using the matrix solid-phase dispersion technique. After that, they were silylated and determined by gas chromatography combined to tandem mass spectrometry (GC/MS/MS). The influence of several factors on the yield and selectivity of the extraction was evaluated in detail. Under final working conditions, samples (0.5 g) were mixed with the same amount of anhydrous sodium sulfate and dispersed on 1.25 g of C18. This blend was transferred to the top of a polypropylene cartridge containing 2 g of Florisil. After removing less polar species with 10 mL of dichloromethane, analytes were recovered using 10 mL of acetonitrile. This extract was concentrated to 1 mL, derivatized, and injected in the GC/MS/MS system. Derivatization was carried out at 45 degrees C in 5 min using 100 microL of N-methyl-N-(tert-butyldimethylsilyl) trifluoroacetamide. Quantification limits from 0.6 to 2.6 ng/g and absolute recoveries between 80 and 114% were achieved. Analysis of dust samples demonstrated the presence of the target species in indoor dust from private houses. The highest average concentration (702 ng/g) corresponded to triclosan.

Optimisation of a solid-phase microextraction method for the determination of parabens in water samples at the low ng per litre level.

A procedure for the determination of five esters of p-hydroxybenzoic acid (parabens) in water samples is presented. Analytes (methyl, ethyl, propyl, butyl and benzyl paraben) are concentrated on a solid-phase microextraction (SPME) fibre, converted on their tert-butyldimethylsilyl derivatives and selectively determined using gas chromatography in combination with tandem mass spectrometry (GC-MS/MS). Influence of different factors on the efficiency of extraction and on-fibre derivatization steps is described in detail. For all species, the highest enrichment factors were achieved using a polyacrylate (PA) fibre exposed directly to stirred water samples, containing 150 mg/ml of sodium chloride, at room temperature. Performance of the further on-fibre derivatization reaction was also maximum at room temperature, considering a short exposition period of the SPME fibre to vapours of the silylation reagent. Under optimised conditions, the proposed method achieved quantification limits from 0.001 to 0.025 ng/ml and it was free of matrix effects; therefore, external calibration can be used as the quantification technique. From our knowledge, this work describes the first application of SPME and gas chromatography to the determination of parabens in water. The analysis of a limited number of real samples revealed the presence of parabens in raw sewage water at concentrations up to 3 ng/ml.

Formation of halogenated by-products of parabens in chlorinated water.

Chemical transformations of four alkyl esters of p-hydroxybenzoic acid, parabens, in chlorinated water samples are investigated. Quantification of the parent species and identification of their reaction by-products were performed using gas chromatography in combination with mass spectrometry. Experiments were accomplished considering free chlorine and paraben concentrations at the mg L(-1) and microg L(-1) level, respectively. Concentration of water samples, using solid-phase extraction, and silylation of the target species were carried out in order to improve the detectability of parent species and their possible transformation products, achieving quantification limits at the low ng L(-1) level. Under employed experimental conditions, the decrease in the concentrations of parabens followed pseudo-first-order kinetics. Half-lives values obtained for model ultrapure water solutions were in good agreement with those observed in tap water samples. For the first type of sample, only two by-products were detected for each paraben. They corresponded to chlorination of the aromatic ring in one or two carbons situated in ortho-positions to the hydroxyl group. Both species were also generated after the addition of parabens to chlorinated tap water. Moreover, three new transformation products were noticed for each parent compound. They were identified as bromo- and bromochloro-parabens, formed due to the existence of traces of bromide in tap water sources. Experiments carried out by mixing paraben-containing personal care products with tap water, containing free chlorine, confirmed the formation of all above described halogenated by-products. In addition, the presence of the di-chlorinated forms of methyl and propyl paraben has been detected for first time in raw sewage water samples.

Aquatic degradation of triclosan and formation of toxic chlorophenols in presence of low concentrations of free chlorine.

The degradation of 2-(2,4-dichlorophenoxy)-5-chlorophenol (triclosan) in chlorinated water samples was investigated. Sensitive determination of the parent compound and its transformation products was achieved by gas chromatography with mass spectrometry detection after sample concentration, using a solid-phase extraction sorbent and silylation of the target compounds. Experiments were accomplished using ultrapure water spiked with chlorine and triclosan concentrations in the low mg/l and ng/ml ranges respectively. Chlorination of the phenolic ring and cleavage of the ether bond were identified as the main triclosan degradation pathways. Both processes led to the production of two tetra- and a penta-chlorinated hydroxylated diphenyl ether, as well as 2,4-dichlorophenol. The formation of 2,3,4-trichlorophenol was not detected in any experiment; however, significant amounts of 2,4,6-trichlorophenol were noticed. All of these five compounds were also identified when triclosan was added to tap-water samples with free chlorine concentrations below 1 mg/l. Minor amounts of three di-hydroxylated phenols, containing from one to three atoms of chlorine in their structures, were also identified as unstable triclosan chlorination by-products. The analysis of several raw wastewater samples showed the co-existence of important concentrations of triclosan and its most stable by-products (2,4-dichlorophenol and 2,4,6-trichlorophenol), reinforcing the potential occurrence of the described transformations when products containing triclosan are mixed with chlorinated tap water.

Microwave assisted extraction followed by gas chromatography with tandem mass spectrometry for the determination of triclosan and two related chlorophenols in sludge and sediments.

A procedure for the determination of 2-(2,4-dichlorophenoxy)-5-chlorophenol (Triclosan) and two possible transformation compounds, 2,4-dichlorophenol (2,4-DCP) and 2,4,6-trichlorophenol (2,4,6-TCP), in sludge from sewage treatment plants (STP) and sediments is presented. Extraction was performed using an acetone:methanol (1:1) mixture under the action of a microwave field. The centrifuged supernatant was diluted with a NaOH aqueous solution and twice extracted with n-hexane for removing neutral and basic interferences. The aqueous layer was acidified and processed as a waste water sample. After concentration analytes were silylated and determined by gas chromatography with tandem mass spectrometry (GC-MS/MS). Influence of experimental conditions on the yield of the extraction process and on the efficiency of the further clean-up step was thoroughly evaluated. Performance of MS/MS detection in comparison to single MS is described. Under final working conditions quantification limits between 0.4 and 0.8 ng/g and recoveries from 78% to 106% were obtained. The method was applied to the analysis of several sludge and sediment samples. Only low levels of TCS were detected in some of the sediments; however, all three compounds were found in sludge samples at concentrations ranging from 7 to 316 ng/g, in the case of chlorophenols, and from 420 to 5400 ng/g, for Triclosan.

Optimization of solid-phase microextraction conditions for the determination of triclosan and possible related compounds in water samples.

A solid-phase microextraction (SPME) method for the determination of triclosan, methyl triclosan, 2,4-dichlorophenol and 2,3,4-trichlorophenol (considered as possible triclosan metabolites) in water samples was optimised. Analytes were first concentrated on a SPME fibre, directly exposed to the sample, and then triclosan and the two chlorinated phenols on-fibre silylated using N-methyl-N-(tert.-butyldimethylsilyl)-trifluoroacetamide (MTBSTFA). Methyl triclosan remained unaffected during the derivatization step. Compounds were determined using gas chromatography in combination with mass spectrometry (GC-MS). Influence of different factors on the efficiency of extraction and derivatization steps was systematically investigated. Using a polyacrylate (PA) fibre quantification limits below 10 ng/l, and acceptable relative standard deviations, were obtained for all compounds after an extraction time of 30 min. On-fibre silylation was carried out in only 10 min. Moreover, the efficiency of the procedure was scarcely affected by the type of water sample. The method was applied to several samples of treated and raw wastewater, triclosan was found in all samples, at concentrations from 120 to 14,000 ng/l, and 2,4-dichlorophenol in most of them, at levels up to 2222 ng/l.

[Prevalence and features of headache and migraine in childhood]. / Prevalencia y características de la cefalea y la migraña en la infancia.

INTRODUCTION: A study of the prevalence of childhood cephalea has been carried on. MATERIAL AND METHODS: We studied a sample of children aged from 3 to 14 years old that consulted to different public health services clinics for control routine. We used an interview based in a questionnaire planned to be answered by parents and the child himself. RESULTS AND CONCLUSIONS: The more relevant results were: 1. Prevalence of cephalea 42%; 2. Prevalence of migraine according to Vahlquist and Kurtz criteria: 8.7% and 6.5% respectively (confidence intervals between 4.4% and 11.1%); 3. Higher proportion of girls for both cephalea and migraine; 4. Higher frequency of cephalea and the migraine type headache are facts related to the decision of seeking medical assistance; 5. 87% of children with migraine from the sample have relatives with intermittent cephalea; 6. Patients selected as having migraine, according to Vahlquist criteria, have the following predominant symptoms: Unilateral condition, vomiting/ nausea, improving with sleep, photophobia, phonophobia, changes in physical facial appearance, need to interrupt activities, pulsation quality, and visual aura.