Juliane Elisa Welke

Characterization of the volatile profile of Brazilian Merlot wines through comprehensive two dimensional gas chromatography time-of-flight mass spectrometric detection.

Wine aroma is an important characteristic and may be related to certain specific parameters, such as raw material and production process. The complexity of Merlot wine aroma was considered suitable for comprehensive two-dimensional gas chromatography (GC×GC), as this technique offers superior performance when compared to one-dimensional gas chromatography (1D-GC). The profile of volatile compounds of Merlot wine was, for the first time, qualitatively analyzed by HS-SPME-GC×GC with a time-of-flight mass spectrometric detector (TOFMS), resulting in 179 compounds tentatively identified by comparison of experimental GC×GC retention indices and mass spectra with literature 1D-GC data and 155 compounds tentatively identified only by mass spectra comparison. A set of GC×GC experimental retention indices was also, for the first time, presented for a specific inverse set of columns. Esters were present in higher number (94), followed by alcohols (80), ketones (29), acids (29), aldehydes (23), terpenes (23), lactones (16), furans (14), sulfur compounds (9), phenols (7), pyrroles (5), C13-norisoprenoids (3), and pyrans (2). GC×GC/TOFMS parameters were improved and optimal conditions were: a polar (polyethylene glycol)/medium polar (50% phenyl 50% dimethyl arylene siloxane) column set, oven temperature offset of 10 °C, 7s as modulation period and 1.4s of hot pulse duration. Co-elutions came up to 138 compounds in 1D and some of them were resolved in 2D. Among the co-eluted compounds, thirty-three volatiles co-eluted in both 1D and 2D and their tentative identification was possible only due to spectral deconvolution. Some compounds that might have important contribution to aroma notes were included in these superimposed peaks. Structurally organized distribution of compounds in the 2D space was observed for esters, aldehydes and ketones, alcohols, thiols, lactones, acids and also inside subgroups, as occurred with esters and alcohols. The Fischer Ratio was useful for establishing the analytes responsible for the main differences between Merlot and non-Merlot wines. Differentiation among Merlot wines and wines of other grape varieties were mainly perceived through the following components: ethyl dodecanoate, 1-hexanol, ethyl nonanoate, ethyl hexanoate, ethyl decanoate, dehydro-2-methyl-3(2H)thiophenone, 3-methyl butanoic acid, ethyl tetradecanoate, methyl octanoate, 1,4 butanediol, and 6-methyloctan-1-ol.

Differentiation of wines according to grape variety using multivariate analysis of comprehensive two-dimensional gas chromatography with time-of-flight mass spectrometric detection data

Headspace solid-phase microextraction (HS-SPME) combined with comprehensive two-dimensional gas chromatography with time-of-flight mass spectrometry detection (GC×GC/TOFMS) was used to analyse the volatiles in five types of wines elaborated with grapes of Cabernet Sauvignon, Merlot, Chardonnay, Sauvignon Blanc and Pinot Noir varieties. Fisher ratio, principal component analysis (PCA) and stepwise linear discriminant analysis (SLDA) were used to develop a discriminant model and, as a result, 12 volatile compounds enabled differentiation and classification of wines according to grape cultivars. A detailed examination of GC×GC/TOFMS data showed that the use of one-dimensional gas chromatography with a mass spectrometric detector (1D-GC/MS) would probably result in misidentification of some of these 12 compounds, as they showed partial coelution with other components in the first chromatographic dimension.

Determination of ochratoxin A in wine by high-performance thin-layer chromatography using charged coupled device

A method was developed and validated in-house for determination of ochratoxin A (OTA) in wine using charge coupled device (CCD) to acquire the fluorescence images of mycotoxin from high-performance thin-layer chromatography (HPTLC) under UV lamp. The method showed a mean recovery of 90.4%. The quantification and detection limist were 0.1 μg L -1 and 0.016 μg L -1 per spot, respectively. The results of validation confirmed the efficiency of the method, which is sensitive enough to be used to quantify OTA in wine. The occurrence of OTA in Brazilian wines was evaluated. OTA was found in one sample at a level of 4.5 μg L -1 , which is higher than the limit of 2 μg L -1 considered acceptable by the Scientific Commission of the European Communities. This study demonstrated the applicability of HPTLC using CCD as a tool to determine OTA in wine.

Volatile characterization by multivariate optimization of headspace-solid phase microextraction and sensorial evaluation of chardonnay base wines

Very little is known about the aroma of wines produced in the South part of Brazil, a region that is responsible for 90% of the Brazilian wine production. A headspace solid phase micro extraction (HS-SPME) method was developed for the extraction of volatile and semivolatile compounds related to the aroma of base wines of five different wineries of the state of Rio Grande do Sul (Brazil). The relationships among volatile composition, aroma quality and sensory evaluation of Chardonnay base wines were performed in this study. Optimum conditions for HS-SPME included use of DVB‑CAR‑PDMS coating, wine samples without further dilution, no stirring, sample volume of 1 mL, 30% of NaCl, extraction time and temperature of 45 min and 55 oC, respectively. Eighteen compounds were identified and fourteen were tentatively identified in the samples. The majority (56%) of them was esters and other minor components were alcohols and acids. The sensory analysis of Vale dos Vinhedos and Pinto Bandeira samples showed the highest quality scores, characterized by higher concentrations of ethyl esters, which contributes to the fruity aroma of these wines.

Quantitative analysis of patulin in apple juice by thin-layer chromatography using a charge coupled device detector.

A method was developed and validated in-house for the detection and quantification of patulin in apple juice concentrate using a charge coupled device (CCD) on thin-layer chromatography (TLC) plates. Samples were extracted with ethyl acetate and then cleaned-up by extraction with a sodium carbonate solution. The method showed a mean recovery of 95%. The quantification and detection limit were 14 µg l−1 and 0.005 µg per spot, respectively. The CCD camera is sufficiently sensitive to detect changes in spot fluorescence intensity caused by small differences in mycotoxin concentration under homogeneous illumination from a UV light source. The results of validation confirmed the efficiency of the method, which is sensitive enough to be used to quantify patulin in apple juice by producers or for government monitoring/survey programs. The method was applied to the analysis of 16 apple juice concentrate samples and patulin levels ranged from 15 to 46 µg l−1. This study demonstrated the applicability of the TLC–CCD technique as a tool for monitoring patulin in apple juice.

Main differences between volatiles of sparkling and base wines accessed through comprehensive two dimensional gas chromatography with time-of-flight mass spectrometric detection and chemometric tools

The main changes in the volatile profile of base wines and their corresponding sparkling wines produced by traditional method were evaluated and investigated for the first time using headspace solid-phase microextraction combined with comprehensive two-dimensional gas chromatography with time-of-flight mass spectrometry detection (GC×GC/TOFMS) and chemometric tools. Fisher ratios helped to find the 119 analytes that were responsible for the main differences between base and sparkling wines and principal component analysis explained 93.1% of the total variance related to the selected 78 compounds. It was also possible to observe five subclusters in base wines and four subclusters in sparkling wines samples through hierarchical cluster analysis, which seemed to have an organised distribution according to the regions where the wines came from. Twenty of the most important volatile compounds co-eluted with other components and separation of some of them was possible due to GC×GC/TOFMS performance.

Comprehensive Two-Dimensional Gas Chromatography for Analysis of Volatile Compounds in Foods and Beverages

A cromatografia gasosa bidimensional abrangente (GC×GC) e uma tecnica analitica relativamente nova, que tem sido amplamente empregada para analise de varios tipos de amostras complexas. A GC×GC proporciona maior capacidade de pico, sensibilidade e resolucao, quando comparada a cromatografia gasosa monodimensional (1D-GC). Alem disso, a estruturacao dos cromatogramas bidimensionais auxilia na identificacao de classes de compostos. No que diz respeito a analise de alimentos e bebidas, a eficiencia superior da GC×GC resulta em beneficios unicos quando comparada as tecnicas analiticas disponiveis para os mais variados objetivos de analise: separacao de grupos de compostos aroma-ativos, verificacao de perfil cromatografico, analise de compostos alvo ou identificacao de componentes desconhecidos. Quando compostos aroma-ativos estao presentes em nivel de tracos, a GC×GC tambem pode ser empregada como uma tecnica de alto potencial devido a sua sensibilidade superior. Tendo em vista que a fracao volatil de muitos alimentos e bebidas pode determinar a aceitacao ou rejeicao destes produtos por parte do consumidor, a caracterizacao da mesma pode ser feita com base na composicao destes compostos volateis. Neste trabalho, as aplicacoes da GC×GC para analise de volateis em alimentos e bebidas sao apresentadas, desde a fase inicial de sua implementacao ate 2010. Alguns topicos relacionados aos principios da GC×GC foram tambem discutidos.

Patulin accumulation in apples during storage by Penicillium expansum and Penicillium griseofulvum strains

A part of apples destined to juice production is generally of poor quality. Apples from cold storage or recently harvest (ground harvested or low quality apples) are stored under ambient conditions until they are processed. Since Penicillium expansum and P. griseofulvum are the principal fungal species isolated from stored apples in Brazil, the objective of this study was to investigate the ability of these strains to produce patulin in apples and report the consequences of this type of storage in loss of quality. The toxin was quantified using thin layer chromatography and charge-coupled device camera (TLC-CCD). The rate and quantities that P. expansum and P. griseofulvum can grow and produce patulin are highly dependent on the fungal strain and time. Lesion diameter resulted to be independent of the strain considered. The maximum period of time which apples were kept at cold storage (4 °C) without patulin accumulation was 27 days. When these apples were kept at 25 °C during 3 days, both factors lesion diameter and patulin production increased significantly. These results confirm that time in which apples are taken out from cold storage room before juice production is critical in order to prevent patulin accumulation.

Monitoring the evolution of volatile compounds using gas chromatography during the stages of production of Moscatel sparkling wine

This study reports, for the first time, the main changes that occur with some important aroma compounds of Moscatel sparkling wines during winemaking, measured using headspace solid-phase microextraction, one-dimensional and comprehensive two-dimensional gas chromatography (GC×GC) with mass spectrometry detection (MS). The best conditions of volatile extraction included the use of PDMS/DVB fibre, 2mL of wine, 30% of NaCl, 40°C for 30min. The chromatographic profile of sparkling wines showed decreasing amounts of monoterpenes (limonene, 4-terpineol, terpinolene, citronellol, α-terpineol, linalool, hotrienol, and nerol oxide), increasing amounts of esters (terpenyl esters, ethyl octanoate, ethyl decanoate and hexyl acetate) and alcohols (1-nonanol and 2-phenylethanol). Sixty-nine compounds co-eluted in the first dimension; only six co-eluted in the second dimension. GC×GC/TOFMS allows more detailed study of the volatile profile of sparkling wines.

Photometric procedure for quantitative analysis of Aflatoxin B1 in peanuts by thin-layer chromatography using charge coupled device detector

A photometric procedure was developed for determination of aflatoxin B1 in peanuts by TLC-CCD technique. The quantification and detection limit were 1.2 μg kg-1 and 0.4 ng per spot, respectively, with mean recovery of 98%. The CCD camera is sufficiently sensitive to detect small changes in spots fluorescence intensity and the results for performance confirmed the efficiency of the method. Another important property of CCD detector is its linearity for a wide range of luminous stimulus determined by analysis of five-point calibration curves using the intensity of AFB1 fluorescence versus AFB1 concentration (0.8 to 4.8 ng per spot). The method was applied to the analysis of thirty nine peanut samples and aflatoxin B1 levels ranged from 16 to 115 μg kg-1. The TLC-CCD and the photometric procedure developed in this study demonstrated to be a simple and efficient tool for quantitative analyses of AFB1 in peanut samples.