San Juan de la Rambla, Spain
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Herrero M.,Institute of Industrial Fermentations CSIC | Herrero M.,Autonomous University of Madrid | Simo C.,Institute of Industrial Fermentations CSIC | Garcia-Canas V.,Institute of Industrial Fermentations CSIC | And 2 more authors.
Electrophoresis | Year: 2010

This review article addresses the different chiral capillary electrophoretic methods used to study and characterize foods and beverages through the enantiomeric separation of different food compounds such as amino acids, pesticides, polyphenols, etc. This work intends to provide an updated overview on the main applications of such enantioselective procedures together with their main advantages and drawbacks in food analysis. Some foreseeable applications and developments of these chiral CZE, CEC and MEKC methods for food characterization are also discussed. Papers that were published within the period January 2003 to October 2009 are included, following the previous review on this topic by Simo et al. (Electrophoresis 2003, 24, 2431-2441). © 2010 Wiley-VCH Verlag GmbH & Co. KGaA.


Manzano P.,University of Valladolid | Arnaiz E.,University of Valladolid | Diego J.C.,University of Valladolid | Toribio L.,University of Valladolid | And 4 more authors.
Journal of Chromatography A | Year: 2011

A method to separate FAME and the linoleic and linolenic acids isomers by GCxGC using an apparatus equipped with a capillary flow technology (CFT) based modulator and a FID detector has been developed. Four different column combinations (one conventional and three inverted phase sets) were used in these experiments. The conventional set first involved a DB5-MS non-polar column followed by a highly polar HP-INNOWax column in the second dimension. The inverted phase set comprised of a highly polar BPX-70 column in the first dimension and a non-polar ZB5-MS column for the second dimension. Furthermore, the influence of the length of the second dimension column on FAME isomer separation was studied in the inverted phase sets, along with other parameters like the modulation time and column flow. The best results in terms of the time required for the analysis and number of FAME identified with the inverted set were achieved with the shorter second dimension column. After supercritical fluid extraction, the method was applied to identify FAMEs in broccoli leaves from three different cultivars (Naxos, Nubia and Viola). © 2011 Elsevier B.V.


Klejdus B.,Mendel University in Brno | Lojkova L.,Mendel University in Brno | Plaza M.,Institute of Industrial Fermentations CSIC | Snoblova M.,Mendel University in Brno | Sterbova D.,Mendel University in Brno
Journal of Chromatography A | Year: 2010

New hyphenated technique for the extraction and determination of isoflavones in sea and freshwater algae and cyanobacteria was developed. The method consists of sonication sample pretreatment, extraction by supercritical CO2 modified by 3% (v/v) of MeOH/H2O mixture (9:1, v/v) at 35MPa and 40°C for 60min, fast chromatography analysis by the means of Agilent 1200 Series Rapid Resolution and MS/MS determination. Agilent 1200 Series RRLC was used with Zorbax SB-CN chromatographic column (100mm×2.1mm, particle size 3.5μm), 3μl injection volume, mobile phase consisting of 0.2% (v/v) acetic acid in water (solvent A) and acetonitrile (solvent B) and used with linear gradient (30% B at 0min, from 0min to 3min up to 50% B, from 3 to 6min up to 80% B and from 6 to 10min down to 30% B). The flow-rate was 0.4mL/min, column oven temperature 35°C. MS detector Agilent Technologies 6460 Triple quadrupole LC/MS with Agilent Jet Stream was used in a negative ESI mode under following conditions: gas temperature 350°C, gas flow 13L/min, nebulizer gas pressure 50psi, sheath gas temperature 400°C, sheath gas flow 12L/min, capillary voltage was 4kV. Samples were analysed in the multiple reaction monitoring (MRM) mode. Eight isoflavone compounds were found for the first time in seven real samples of sea algae and in three control samples of freshwater algae and cyanobacteria. Usual optimisation study of extraction parameters was performed. Pressure and temperature optima for algae matrix are different from those obtained sooner for other matrices for most of the analytes, but the results of modifier optimisation study are in good accordance with those obtained sooner for spiked samples and red clover matrix. It seems that matrix has very small or no effect on the modifier selection. Two different approaches of sonication pretreatment were tested: sonication bath and the thorn instrument. In longer extraction time experiments, thorn sonication was more efficient and recovery of following supercritical fluid extraction was higher. © 2010 Elsevier B.V.


Ongay S.,Institute of Organic Chemistry CSIC | Martin-Alvarez P.J.,Institute of Industrial Fermentations CSIC | Neusu C.,Aalen University of Applied Sciences | de Frutos M.,Institute of Organic Chemistry CSIC
Electrophoresis | Year: 2010

α-1-Acid glycoprotein (AGP) is a highly heterogeneous protein that presents a vast number of isoforms (molecules of the protein differing in its peptidic and/or glycosidic moieties). In recent years, several authors have studied the potential use of AGP as a cancer biomarker. These studies focus on the correlation of different features of AGP structure (i.e. fucosylation, antennarity) with cancer or on the total protein blood concentration. In this study, the potential of CZE-UV and CZE-ESI-MS analysis of intact AGP isoforms to study the correlation of this protein with bladder cancer is shown. Samples from 16 individuals (eight healthy, eight bladder cancer) were analyzed and characterized in great detail including data on intact protein isoforms and on released glycans. The analytical data were evaluated employing different statistical techniques (ANOVA; principal component analysis, linear discriminant analysis; and partial least squares-discriminant analysis). Statistical differences between the two groups of study were observed. The best results were obtained by linear discriminant analysis of the CZEESI-MS data for intact AGP isoforms (93.75% of correct classification). Due to MS characterization, it can be observed that differences between the samples are mainly due to higher abundance of AGP isoforms containing tri- and tetra-antennary fucosylated oligosaccharides in cancer patients. The results show the great potential of CE-MS in combination with advanced data processing for the use of intact protein isoforms as disease biomarkers. © 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.


Monagas M.,Institute of Industrial Fermentations CSIC | Urpi-Sarda M.,University of Barcelona | Sanchez-Patan F.,Institute of Industrial Fermentations CSIC | Llorach R.,University of Barcelona | And 4 more authors.
Food and Function | Year: 2010

Flavan-3-ols, occurring in monomeric, as well as in oligomeric and polymeric forms (also known as condensed tannins or proanthocyanidins), are among the most abundant and bioactive dietary polyphenols, but their in vivo health effects in humans may be limited because of their recognition as xenobiotics. Bioavailability of flavan-3-ols is largely influenced by their degree of polymerization; while monomers are readily absorbed in the small intestine, oligomers and polymers need to be biotransformed by the colonic microbiota before absorption. Therefore, phenolic metabolites, rather than the original high molecular weight compounds found in foods, may be responsible for the health effects derived from flavan-3-ol consumption. Flavan-3-ol phenolic metabolites differ in structure, amount and excretion site. Phase II or tissular metabolites derived from the small intestine and hepatic metabolism are presented as conjugated derivatives (glucuronic acid or sulfate esters, methyl ether, or their combined forms) of monomeric flavan-3-ols and are preferentially eliminated in the bile, whereas microbial metabolites are rather simple conjugated lactones and phenolic acids that are largely excreted in urine. Although the colon is seen as an important organ for the metabolism of flavan-3-ols, the microbial catabolic pathways of these compounds are still under consideration, partly due to the lack of identification of bacteria with such capacity. Studies performed with synthesized or isolated phase II conjugated metabolites have revealed that they could have an effect beyond their antioxidant properties, by interacting with signalling pathways implicated in important processes involved in the development of diseases, among other bioactivities. However, the biological properties of microbe-derived metabolites in their actual conjugated forms remain largely unknown. Currently, there is an increasing interest in their effects on intestinal infections, inflammatory intestinal diseases and overall gut health. The present review will give an insight into the metabolism and microbial biotransformation of flavan-3-ols, including tentative catabolic pathways and aspects related to the identification of bacteria with the ability to catabolize these kinds of polyphenols. Also, the in vitro bioactivities of phase II and microbial phenolic metabolites will be covered in detail. © The Royal Society of Chemistry.


PubMed | National Research Council Italy and Institute of Industrial Fermentations CSIC
Type: Journal Article | Journal: Food chemistry | Year: 2015

Determination of amino acid enantiomers is a very important topic in food analysis, since the presence of d-isomers may indicate, e.g., adulteration, microbiological contamination, uncontrolled fermentation processes, etc. In fact, the d- and l-enantiomers contents can be a useful marker for several elements such as quality control, contamination detection, processing monitoring, etc. Here we studied the potentiality of nano-liquid chromatography (nano-LC) coupled with mass spectrometry for the enantiomeric separation of several d- and l-amino acids that can be found in food products. Analytes were derivatized with fluorescein isothiocyanate (FITC). The mixture was injected and compounds focused on a C18 cartridge, then nano-LC analysis was carried out in a capillary column (75m i.d.) packed with vancomycin-modified silica-diol particles. The effect of some experimental parameters, such as pH and buffer concentration on enantioresolution and retention factors, was studied for method optimization. The chromatographic separation system was coupled with an ion-trap mass spectrometer through a nano spray interface. It provided a final evaluation on analytes detected in all investigated samples with LOD values as low as 8ng/mL. That method was applied to the comparative analysis of two different orange juice samples (fresh natural vs. commercial one). Obtained profiles confirmed expected high quality standards. In fact, they mainly contained l-amino acids forms and not their antipodes.


Garcia-Canas V.,Institute of Industrial Fermentations CSIC | Simo C.,Institute of Industrial Fermentations CSIC | Leon C.,Institute of Industrial Fermentations CSIC | Ibanez E.,Institute of Industrial Fermentations CSIC | And 2 more authors.
Mass Spectrometry Reviews | Year: 2011

The development of genetically modified crops has had a great impact on the agriculture and food industries. However, the development of any genetically modified organism (GMO) requires the application of analytical procedures to confirm the equivalence of the GMO compared to its isogenic non-transgenic counterpart. Moreover, the use of GMOs in foods and agriculture faces numerous criticisms from consumers and ecological organizations that have led some countries to regulate their production, growth, and commercialization. These regulations have brought about the need of new and more powerful analytical methods to face the complexity of this topic. In this regard, MS-based technologies are increasingly used for GMOs analysis to provide very useful information on GMO composition (e.g., metabolites, proteins). This review focuses on the MS-based analytical methodologies used to characterize genetically modified crops (also called transgenic crops). First, an overview on genetically modified crops development is provided, together with the main difficulties of their analysis. Next, the different MS-based analytical approaches applied to characterize GM crops are critically discussed, and include "-omics" approaches and target-based approaches. These methodologies allow the study of intended and unintended effects that result from the genetic transformation. This information is considered to be essential to corroborate (or not) the equivalence of the GM crop with its isogenic non-transgenic counterpart. © 2010 Wiley Periodicals, Inc.


Garcia-Canas V.,Institute of Industrial Fermentations CSIC | Simo C.,Institute of Industrial Fermentations CSIC | Leon C.,Institute of Industrial Fermentations CSIC | Cifuentes A.,Institute of Industrial Fermentations CSIC
Journal of Pharmaceutical and Biomedical Analysis | Year: 2010

In recent years, nutrition research has moved from classical epidemiology and physiology to molecular biology and genetics. Following this trend, Nutrigenomics has emerged as a novel and multidisciplinary research field in nutritional science that aims to elucidate how diet can influence human health. It is already well known that bioactive food compounds can interact with genes affecting transcription factors, protein expression and metabolite production. The study of these complex interactions requires the development of advanced analytical approaches combined with bioinformatics. Thus, to carry out these studies Transcriptomics, Proteomics and Metabolomics approaches are employed together with an adequate integration of the information that they provide. In this article, an overview of the current methodologies and a thorough revision of the advances in analytical technologies and their possibilities for future developments and applications in the field of Nutrigenomics is provided. © 2009 Elsevier B.V. All rights reserved.


Simo C.,Institute of Industrial Fermentations CSIC | Garcia-Canas V.,Institute of Industrial Fermentations CSIC | Cifuentes A.,Institute of Industrial Fermentations CSIC
Electrophoresis | Year: 2010

This review article addresses the developments and applications of capillary electromigration methods coupled on-line with MS for chiral analysis. The multiple enantiomeric applications of this hyphenated technology are covered including chiral analysis of drugs, food compounds, pesticides, natural metabolites, etc. in different matrices such as plasma, urine, medicines, foods, etc. This work intends to provide an updated overview (including works published till September 2009) on the principal chiral applications carried out by CZE-MS, CEC-MS and MEKC-MS, discussing their main advantages and drawbacks in all their different areas of application as well as their foreseeable development in the not too distant future. © 2010 Wiley-VCH Verlag GmbH & Co. KGaA.


Garcia-Canas V.,Institute of Industrial Fermentations CSIC | Mondello M.,Institute of Industrial Fermentations CSIC | Cifuentes A.,Institute of Industrial Fermentations CSIC
Electrophoresis | Year: 2010

In this work, an innovative method useful to simultaneously analyze multiple genetically modified organisms is described. The developed method consists in the combination of multiplex ligation-dependent genome dependent amplification (MLGA) with CGE and LIF detection using bare-fused silica capillaries. The MLGA process is based on oligo-nucleotide constructs, formed by a universal sequence (vector) and long specific oligo-nucleotides (selectors) that facilitate the circularization of specific DNA target regions. Subsequently, the circularized target sequences are simultaneously amplified with the same couple of primers and analyzed by CGE-LIF using a bare-fused silica capillary and a run electrolyte containing 2-hydroxyethyl cellulose acting as both sieving matrix and dynamic capillary coating. CGE-LIF is shown to be very useful and informative for optimizing MLGA parameters such as annealing temperature, number of ligation cycles, and selector probes concentration. We demonstrate the specificity of the method in detecting the presence of transgenic DNA in certified reference and raw commercial samples. The method developed is sensitive and allows the simultaneous detection in a single run of percentages of transgenic maize as low as 1% of GA21, 1% of MON863, and 1% of MON810 in maize samples with signal-to-noise ratios for the corresponding DNA peaks of 15, 12, and 26, respectively. These results demonstrate, to our knowledge for the first time, the great possibilities of MLGA techniques for genetically modified organisms analysis. © 2010 Wiley-VCH Verlag GmbH & Co. KGaA.

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