Extraction of wood compounds by use of subcritical fluids

A study of the extraction of oak wood compounds with subcritical water-ethanol mixtures as extractant, with an ethanol content between 0-60%, is reported. Identification and characterisation of the extracted compounds have been made by spectrophotometry and gas chromatography with either flame ionisation or mass detectors. Extraction was performed statically manner by use of a single cycle or repeated cycles. All variables affecting the extraction process were studied and optimised. Extraction time and temperature are 60 min and 200ºC, respectively. Comparison of the extract thus obtained with commercial extracts showed the former to be rich in compounds characteristic of the commercial extracts. The method enables manipulation of the extract composition by changing the temperature and water/ethanol ratio used. It is faster than the traditional procedures for obtaining wood extracts

Two-parameter determination in vinegar by a flow injection-pervaporation system

A flow-injection method (FI) for the sequential determination of ethanol and acetic acid in vinegar is reported. The determination of ethanol is based on the oxidation of the pervaporated ethanol by Cr2O7K2. The acetic acid is determined by an acid-base reaction with Thymol Blue as the indicator. Both reactions are monitored photometrically at 600 nm using a single detector. Optimisation studies and assessment of the sequential FI method are also reported. The linear determination range is between 0-12 % (v/v) for ethanol and 0-10 % (grams of acetic acid in 100 ml) for acetic acid. The sample throughput of the sequential manifold is 7 per hour. The new method has been applied to vinegar samples and the results obtained are in excellent agreement with those from the reference methods used in Spain

Semiautomatic flow-injection method for the determination of volatile acidity in wines

A flow-injection (FI) method based on analytical pervaporation was assessed for its routine use in the determination of volatile acidity in winery laboratories. The new method was compared with both the official method and the Mathieu method, which is most often used in Spanish wineries, by testing 30 different wines, including young and aged, and sweet and dry wines from Montilla-Moriles appellation d´origine. The robustness of the new method was established and then all three methods were studied in terms of range of linearity and regression of the calibration curve, repeatability, reproducibility, sensitivity, detection and quantification limits (LOD and LQ, respectively) and time of analysis. The FI method surpasses the Mathieu method in reproducibility and both the Mathieu and official methods in LOD and LQ and sensitivity; it also requires less personnel involvement and shorter analysis time. The statistical criteria established by the “Office International de la Vigne et du Vin” were applied to the data and the results obtained indicated that the differences between the analytical parameters of the 3 methods are not significant and can be applied indistinctly. The correlation of the methods was studied by taking them 2 by 2, and the corresponding equations, coefficients and deviations confirmed the statistical results. Thus, the new method can be used in winery laboratories with clear advantages over its 2 counterparts (the routine and official methods)

Sequential Spectrophotometric determination of methanol and iron in vinegar by a flow injection-pervaporation method

An easily automatable sequential flow-injection-pervaporation method is proposed for the photometric determination of methanol and iron in vinegar. The method is based on separation of the methanol from the sample matrix by pervaporation followed by its oxidation to formaldehyde with permanganate, decolouration of the latter with S2O5-2 and subsequent reaction of formaldehyde with p-rosaniline to yield a violet reaction product with maximum absorption at 567 nm. Iron is determined by an existing method based on reaction with thiocyanate in acidic medium and monitoring at 508 nm. After optimisation by either the univariate or multivariate approaches, as required, the linear range was established for methanol (4-1000 mg L-1) and iron (0.18-20 mg L-1); The proposed method was then compared with reference methods for methanol and iron in terms of repeatability (2.452 mg L-1 and 0.245 mg L-1, respectively), reproducibility (4.435 mg L-1 and 0.356 mg L-1, respectively), detection limit (LOD=82 and 0.234 mg L-1, respectively) and traceability. The throughput was nine samples per hour

Method for the simultaneous determination of total polyphenol and anthocyan indexes in red wines using a flow injection approach

A simultaneous and fast method for the determination of total polyphenol index (t.p.i.) and total anthocyan index (t.a.i.) has been developed by a flow injection approach and a diode array spectrophotometer for monitoring at 280 nm and 520 nm, respectively. Linear ranges were obtained from 20 to 70 index units and from 20 to 500 mg l-1 for t.p.i. and t.a.i., respectively. The results provided by the proposed method agree with those obtained using the polyphenol index at 280 nm and the Ribereau-Gayon method for the determination of total anthocyans. The sample throughput was 25-30 samples per hour. Analytical features such as repeatability, reproducibility and detection and quantification limits as well as the results of a robustness study based on the Steiner-Younden procedure are also given

Determination of ethanol in beverages by flow injection, pervaporation and density measurements

A fast, clean and easy to automate flow injection-pervaporation method for the determination of ethanol in different beverages using density measurements is proposed. The method is based on separation of the ethanol from the sample using a pervaporation module, thus obtaining in the acceptor chamber of the pervaporator a water-alcohol mixture, the density of which is measured. After optimisation by either the univariate or multivariate approach as required, a linear range between 0-40% was established. Then, the assessment of the method versus a reference one was studied in terms of repeatability (0.12% v/v), reproducibility (0.32% v/v), detection limit (0.11% v/v) and traceability. The sample throughput was 15 samples h-1. The method was in agreement with the reference methods used in the European Union

Superheated liquids for the extraction of solid residues from winemaking processes

Solid residues from winemaking process have been subjected to extraction with superheated water-ethanol mixtures. Identification and characterisation of the extracted compounds have been made by spectrophotometry, gas chromatography with either flame ionisation or mass detectors and high performance liquid chromatography with UV detection. The extraction has been performed in a static manner by single or repetitive cycles. All variables affecting the extraction process have been studied and optimised. The extraction time and temperature are 65 min and 210ºC, respectively. Two phases constitute the extract: an aqueous phase, which is rich in phenol compounds and an oily phase, mainly formed by fatty acids. The method allows manipulation of the extract composition by changing the applied pressure, temperature, water-ethanol ratio and pH. The method is faster than the traditional extraction procedures for obtaining valuable compounds from these residues

De novo analysis of the haustorial transcriptome of the cucurbit powdery mildew fungus Podosphaera xanthii reveals new candidate secreted effector proteins

Backgrounds. Cucurbit crops are affected, among other pathogens, by the obligate biotrophic fungus Podosphaera xanthii, the main causal agent of powdery mildew in cucurbits. This fungus develops a specialized structure of parasitism termed haustorium. Haustoria are developed into epidermal cells and are responsible for nutrients uptake and effectors delivery. Objectives. The aim of this study was to obtain the haustorial transcriptome of P. xanthii to complete the panel of effector candidates of this fungal pathogen. Methods. To obtain the haustorial transcriptome, we have developed an effective method for isolation of haustoria without contaminants by flow cytometry. The cDNA library was built using a combination of dT primers and random primers followed by a depletion of ribosomal sequences. Sequencing was carried out by Illumina NextSeq550. Conclusions. After bioinformatic analysis, we were able to identify 25 new effector candidates secreted by the classic pathway (with signal peptide) and 269 new candidates secreted by the non-classic pathway (without signal peptide). Most proteins had no functional annotation. By protein modelling and ligand predictions, we are now being able to assign putative functions to some of these candidates to select those with potential roles in pathogenesis for subsequent functional in vivo analysis by HIGS (host-induced gene silencing). By these approaches, we are starting to shed some light into the molecular mechanisms of pathogenesis in this very important pathogen of cucurbits.This work was supported by a grant from the Ministerio de Economía y Competitividad (AGL2013-41938-R), co-financed with FEDER funds (EU). A grant form Universidad de Málaga, Campus de Excelencia Internacional Andalucía Tech, is also acknowledged