Maria Luisa Ruiz del Castillo

Determination of priority phenolic compounds in water and industrial effluents by polymeric liquid-solid extraction cartridges using automated sample preparation with extraction columns and liquid chromatography use of liquid-solid extraction cartridges for stabilization of phenols

Abstract Fourteen phenolic compounds: catechol, phenol, 4-nitrophenol, 4-methylphenol, 2,4-dinitrophenol, 2-nitrophenol, 2-chlorophenol, 4-chlorophenol, 3-chlorophenol, 4-chloro-3-methylphenol, 2,4-dimethylphenol, 2,4-dichlorophenol, 2,4,6,-trichlorophenol and pentachlorophenol, which are included in the priority pollutants list of the US Environmental Protection Agency and in the European Union list 76/464, were determined in water by liquid-solid extraction (LSE) followed by liquid chromatography with UV detection (LC-UV). Three different polymeric sorbents were used: Isolute ENV+, Lichrolut EN and Porapak RDX. The developed method involves the use of automated sample preparation with extraction columns (ASPEC XL) for automated sample preconcentration and a Baker LSE 12G apparatus with the vacuum set at 15 p.s.i. (1p.s.i.=6894.76Pa) for the drying step. The recoveries varied from 70 to 100% (except for catechol) on preconcentrating 700–1000 ml of a 5 μg/l solution, pH 2.5−4. The stability of the target compounds on Isolute ENV+ was evaluated by storing the sorbent material at −20°C, 4°C and at room temperature for up to three months. Complete recovery was observed after storage at −20°C for two months. At room temperature, losses of up to 70% were observed for phenol, catechol and the more volatile phenols. The stability of the phenolic componds was dependent on the water matrix, the storage temperature and on physico chemical properties, such as vapor pressure and water solubility.

Impact of postharvest methyl jasmonate treatment on the volatile composition and flavonol content of strawberries

BACKGROUND Although strawberry aroma is very complex, certain compounds have been described as main contributors, i.e. furanones, aldehydes, alcohols, sulfur compounds and particularly methyl and ethyl esters. In addition, strawberries possess potent antioxidant activity because of their high content of phenolic compounds. Among them, flavonols are highlighted as important antioxidant compounds in strawberry. The objective of this study was to assess the effect of methyl jasmonate (MJ) on the composition of the major contributors to aroma and on the content of certain flavonols in strawberry fruits. RESULTS The levels of all studied volatile compounds were significantly affected by MJ treatment, though the individual effect differed according to the specific compound considered. Most of them increased significantly (P < 0.05), except methyl butanoate, which always showed higher levels in untreated strawberries. In contrast to aroma compounds, the change in the concentration of flavonols (i.e. myricetin, quercetin and kaempferol) was not significant in MJ-treated strawberries. Considering the health-promoting activity of these compounds, further investigations on the experimental conditions related to the treatment are required to control flavonol bioformation by means of MJ. CONCLUSION The exogenous application of MJ vapour to strawberry enhances, in general, the production of the most relevant aroma-active compounds. On the contrary, MJ treatment does not appear to influence the levels of myricetin, quercetin and kaempferol. Thus postharvest MJ treatment is proposed as an approach to obtain improved strawberry fruits in terms of sensory quality and health-promoting properties.

(+)-methyl jasmonate-induced bioformation of myricetin, quercetin and kaempferol in red raspberries.

The effect of postharvest treatment with enantiomers of methyl jasmonate (MJ) in conjunction with ethanol on bioformation of myricetin, quercetin and kaempferol in red raspberry was studied. For comparison, postharvest treatment with the commercial stereoisomeric mixture of MJ in conjunction with ethanol was simultaneously accomplished. The levels obtained were contrasted with those determined in untreated (control) samples. Exogenous (+)-MJ induced an enhancement in the levels of myricetin, quercetin and, particularly, kaempferol whereas the exposition to (-)-MJ exhibited the opposite effect. Enzymatic assays were carried out in presence and absence of (-)-MJ and (+)-MJ to evaluate possible changes in the activity of the enzymes regulating the bioformation of flavonols in red raspberries as a consequence of the treatments. From the results of the assays both (-)-MJ and (+)-MJ inhibited the activity of flavanone 3β-hydroxylase (FHT) and flavonol synthase (FLS), which are directly involved in the formation of flavonols from (-/+)-naringenin. From these results, it is speculated that the activity of phenylalanine ammonia lyase (PAL) regulating the formation of (-/+)-naringenin from l-phenylalanine by (+)-MJ in conjunction with ethanol is promoted. Postharvest treatment of red raspberry with (+)-MJ in ethanol is proposed as a mean to increase flavonol content in red raspberries.

Evaluation of pesticide residue contents in fruit juice by solid-phase microextraction and multidimensional gas chromatography coupled with mass spectrometry

A new method based on the use of SPME followed by the MDGC-MS analysis was developed to determine pesticides in fruit juice. Different pesticide mixture standards (i.e. Mix 101, 13 and 164) were initially analyzed to optimize the separation conditions. To evaluate the advantages of the two-dimensional system over monodimensional GC, a comparative study on relative standard deviations, detection limits and correlation coefficients was carried out. As a result, selective transfers of some pesticides from the first to the second dimension were at times essential to avoid overlapping. The selected separation conditions from the study with standards were applied to fruit juices spiked with some pesticide standards. The results found in this work prove that the employment of a multidimensional analysis technique permits to avoid false positives obtained frequently on monodimensional techniques as a consequence of interferences of the analytes with matrix components.

Structure and Dynamics of 1-Butyl-3-methylimidazolium Hexafluorophosphate Phases on Silica and Laponite Clay: From Liquid to Solid Behavior

Solid-state NMR experiments show that the behavior of supported 1-butyl-3-methylimidazolium hexafluorophosphate ionic liquid phases depends on the type of support and the phase thickness. A mobile nearly liquid phase is obtained on silica, on the basis of the line widths of the bands in (1)H, (31)P, and (13)C spectra. However, the mobility is somehow restricted, as shown by the possibility of using the cross-polarization technique, although with slow dynamics. On laponite clay, a layered material with a negatively charged surface, a truly solid phase is obtained at low coverage, whereas the increase in ionic liquid loading leads to a second liquid phase, as shown by the presence of two contributions with very different line widths. These two phases seem to coexist without exchange in the NMR time frame. Heteronuclear correlation experiments evidence different relative dispositions of the imidazolium-surface-PF(6) system, with only aromatic protons involved in all the interactions on silica but participation of the benzylic groups (N-CH(3) and/or N-CH(2)) in the case of laponite clay.

Enantioselective isolation of methyl jasmonate using permethyl-β-cyclodextrin HPLC

A method based on the use of HPLC for the enantioselective resolution of the four stereoisomers of methyl jasmonate (MJ) with no need for the previous formation of the diastereoisomers is developed. To that end, a Nucleodex-beta-PM column as well as an optimization process considering different flow rates and mobile phase compositions were required. As a result, 0.8 mL/min and 55:45 methanol/water composition were the conditions selected to carry out the separation of the stereoisomers. Isolation of pure (-)- and (+)-MJ was accomplished by collecting the HPLC fractions corresponding to their elution time. SPE was subsequently used to concentrate and change the solvent of the HPLC fractions collected. Chiral GC and polarimetry were additionally employed to evaluate the purity and optical rotation, respectively, of the enantiomers separated. The results found in this study are particularly relevant considering that MJ stereoisomers are not commercially available.

Development of a method based on on-line reversed phase liquid chromatography and gas chromatography coupled by means of an adsorption-desorption interface for the analysis of selected chiral volatile compounds in methyl jasmonate treated strawberries

A method based on the use of the through oven transfer adsorption-desorption (TOTAD) interface in on-line coupling between reversed phase liquid chromatography and gas chromatography (RPLC-GC) for the determination of chiral volatile compounds was developed. In particular, the method was applied to the study of the influence of methyl jasmonate (MJ) treatment on the production and enantiomeric composition of selected aroma compounds in strawberry. The compounds studied were ethyl 2-methylbutanoate, linalool and 4-hydroxy-2,5-dimethyl-3(2H)-furanone (i.e. furaneol), which were examined on days 3, 6 and 9 after treatment. The method developed resulted in relative standard deviations (RSDs) of 21.6%, 8.1% and 9.8% and limits of detection (LD) of 0.04, 0.07 and 0.02mg/l for ethyl 2-methylbutanoate, linalool and furaneol, respectively. The application of the RPLC-TOTAD-GC method allowed higher levels of ethyl 2-methylbutanoate, linalool and furaneol to be detected, particularly after 9 days of treatment. Besides, MJ demonstrated to affect the enantiomeric distribution of ethyl 2-methylbutanoate. On the contrary, the enantiomeric composition of linalool and furaneol kept constant in both control and MJ-treated strawberries throughout the study. These results are discussed.

Through oven transfer adsorption–desorption interface for the analysis of methyl jasmonate in aromatic samples by on‐line RPLC‐GC

A fully automated method for the determination of medium volatility compounds in aromatic samples was developed. Specifically, the determination of methyl jasmonate in jasmine fragrances was performed by using the through oven transfer adsorption-desorption (TOTAD) interface for the on-line coupling between RPLC-GC. A study of the most relevant variables involved in the performance of the TOTAD interface for medium volatility compounds was carried out by testing different values of helium flow (100, 300, 400, and 500 mL/min), transfer speed (0.1, 0.3, 0.5, and 2.0 mL/min), and methanol/water percentages (86:14, 85:15, 83:17, 80:20, and 70:30). The method developed provided satisfactory repeatability (RSD for retention times of 0.15% and for peak areas of 9.4%) and recovery (71%) as well as excellent LOD (0.01 mg/L) for methyl jasmonate in commercial jasmine essence under the experimental conditions selected as optimum. Additional advantages of the automated RPLC-TOTAD-GC method proposed in the present work are its rapidness, reliability, and the possibility of directly introducing the sample with no further pretreatment.

Online RPLC-GC via TOTAD Method To Isolate (+)-Methyl Epijasmonate from Lemon (Citrus limon Burm.)

Pure (+)-methyl epijasmonate was isolated from lemon ( Citrus limon Burm.) for the first time. To that aim, two commercial essential oils and one homemade extract were included in the present paper. First, a study on the appropriate chromatographic conditions to avoid the epimerization from methyl epijasmonate to the more stable methyl jasmonate was accomplished. The results obtained are discussed. The presence of (+)-methyl epijasmonate in the three samples studied was initially established through the direct injection into GC-MS. However, the overlapping of (+)-methyl epijasmonate with other matrix components made it necessary to employ a multidimensional technique. RPLC-GC analysis via through-oven transfer adsorption-desorption (TOTAD) provided the selectivity and sensitivity required, reflecting that the homemade lemon extract was an adequate natural source to obtain pure (+)-methyl epijasmonate by means of the collection of the corresponding RPLC fraction.

Evaluation of a transfer technique for direct coupling of reversed-phase liquid chromatography with gas chromatography

Abstract A method that allows direct transfer of liquid chromatographic aqueous eluents into capillary gas chromatography, using a programmed temperature vaporizer as interface, was investigated. The method involved both the evaporative and nonevaporative modes of solvent elimination and allowed large fractions of methanol–water eluents to be transferred from LC to GC. The speed of sample transfer was 1800 μl/min and the internal diameter of the column used in the preseparation step was 4.6 mm. Working rules of the method were investigated to improve sensitivity for the direct analysis of trace compounds. Reproducibility was achieved and detection limits ranging from 0.1 to 1.5 μg/ml of ethyl esters covering a wide range of boiling points were obtained.