Anna Vallverdú-Queralt

Improved characterization of tomato polyphenols using liquid chromatography/electrospray ionization linear ion trap quadrupole Orbitrap mass spectrometry and liquid chromatography/electrospray ionization tandem mass spectrometry

Tomato (Lycopersicon esculentum Mill.) is the second most important fruit crop worldwide. Tomatoes are a key component in the Mediterranean diet, which is strongly associated with a reduced risk of chronic degenerative diseases. In this work, we use a combination of mass spectrometry (MS) techniques with negative ion detection, liquid chromatography/electrospray ionization linear ion trap quadrupole-Orbitrap-mass spectrometry (LC/ESI-LTQ-Orbitrap-MS) and liquid chromatography/electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) on a triple quadrupole, for the identification of the constituents of tomato samples. First, we tested for the presence of polyphenolic compounds through generic MS/MS experiments such as neutral loss and precursor ion scans on the triple quadrupole system. Confirmation of the compounds previously identified was accomplished by injection into the high-resolution system (LTQ-Orbitrap) using accurate mass measurements in MS, MS(2) and MS(3) modes. In this way, 38 compounds were identified in tomato samples with very good mass accuracy (<2 mDa), three of them, as far as we know, not previously reported in tomato samples.

A comprehensive study on the phenolic profile of widely used culinary herbs and spices: Rosemary, thyme, oregano, cinnamon, cumin and bay

Herbs and spices have long been used to improve the flavour of food without being considered as nutritionally significant ingredients. However, the bioactive phenolic content of these plant-based products is currently attracting interest. In the present work, liquid chromatography coupled to high-resolution/accurate mass measurement LTQ-Orbitrap mass spectrometry was applied for the comprehensive identification of phenolic constituents of six of the most widely used culinary herbs (rosemary, thyme, oregano and bay) and spices (cinnamon and cumin). In this way, up to 52 compounds were identified in these culinary ingredients, some of them, as far as we know, for the first time. In order to establish the phenolic profiles of the different herbs and spices, accurate quantification of the major phenolics was performed by multiple reaction monitoring in a triple quadrupole mass spectrometer. Multivariate statistical treatment of the results allowed the assessment of distinctive features among the studied herbs and spices.

Comprehensive identification of walnut polyphenols by liquid chromatography coupled to linear ion trap-Orbitrap mass spectrometry.

Epidemiologic studies and clinical trials have demonstrated consistent benefits of walnut consumption on coronary heart disease risk and other chronic diseases. Walnuts (Juglans regia L.) have been described previously as a rich source of polyphenols with a broad array of different structures. However, an accurate screening of its complete phenolic profile is still lacking. In the present work, liquid chromatography coupled with electrospray ionization hybrid linear trap quadrupole-Orbitrap mass spectrometry (LC-LTQ-Orbitrap) was applied for a comprehensive identification of phenolic compounds in walnuts. A total of 120 compounds, including hydrolysable and condensed tannins, flavonoids and phenolic acids were identified or tentatively identified on the base of their retention times, accurate mass measurements and subsequent mass fragmentation data, or by comparing with reference substances and literature. The peak area of each signal in mass chromatograms was used to provide semiquantitative information for comparison purposes. The most abundant ions were observed for ellagitannins, ellagic acid and its derivatives. Furthermore, the high-resolution MS analysis revealed the presence of eight polyphenols that have never been reported in walnuts: stenophyllanin C, malabathrin A, eucalbanin A, cornusiin B, heterophylliin E, pterocarinin B, reginin A and alienanin B.

A comprehensive characterisation of beer polyphenols by high resolution mass spectrometry (LC–ESI-LTQ-Orbitrap-MS)

Beer is the second most consumed alcoholic beverage in Europe and shown by the European Prospective Investigation into Cancer and Nutrition cohort study to be the main food contributor to hydroxybenzoic acid intake. About 70-80% of the total polyphenol content in beer comes from malt, and the remaining 30-20% from hops. In this work, liquid chromatography coupled with an electrospray ionization hybrid linear ion trap quadrupole Orbitrap mass spectrometry technique has been used for an accurate identification of beer polyphenols. 47 phenolic compounds were identified using high mass accuracy and confirmed by MS(2) experiments, including simple phenolic acids, hydroxycinnamoylquinics, flavanols, flavonols, flavones, alkylmethoxyphenols, alpha- and iso-alpha-acids, hydroxyphenylacetic acids and prenylflavonoids. As far as we know, 7 of these compounds have been recognised in beer for the first time: feruloylquinic acid, caffeic acid-O-hexoside, coumaric acid-O-hexoside, sinapic acid-O-hexoside, catechin-O-dihexoside, kaempferol-O-hexoside, and apigenin-C-hexoside-pentoside.

Evaluation of a Method To Characterize the Phenolic Profile of Organic and Conventional Tomatoes

The present study aims to compare the phenolic profiles of organic and conventional tomatoes bought in the market. For the quantification and identification of individual polyphenols, liquid chromatography coupled to mass spectrometry in tandem mode (LC-MS/MS) was carried out. Confirmation of the compounds previously identified on the triple-quadrupole was accomplished by injection in the high-resolution system (QToF-MS). In this way, 34 compounds were identified in tomato fruits. Recoveries of targeted polyphenols exceed 78% for conventional and organic tomatoes, respectively. The method intraday precision ranged between 3 and 5%, whereas the interday one was below 12%. Comparing the analyses of tomatoes from conventional and organic production systems demonstrated statistically higher levels (P < 0.05) of phenolic compounds in organic tomatoes. This methodology allowed finding differences in the bioactive components of organic and conventional tomatoes not previously reported.

Metabolite profiling of phenolic and carotenoid contents in tomatoes after moderate-intensity pulsed electric field treatments.

A metabolite profiling approach was used to study the effect of moderate-intensity pulsed electric field (MIPEF) treatments on the individual polyphenol and carotenoid contents of tomato fruit after refrigeration at 4°C for 24h. The MIPEF processing variables studied were electric field strength (from 0.4 to 2.0kV/cm) and number of pulses (from 5 to 30). Twenty four hours after MIPEF treatments, an increase was observed in hydroxycinnamic acids and flavanones, whereas flavonols, coumaric and ferulic acid-O-glucoside were not affected. Major changes were also observed for carotenoids, except for the 5-cis-lycopene isomer, which remain unchanged after 24h of MIPEF treatments. MIPEF treatments, conducted at 1.2kV/cm and 30 pulses, led to the greatest increases in chlorogenic (152%), caffeic acid-O-glucoside (170%) and caffeic (140%) acids. On the other hand, treatments at 1.2kV/cm and 5 pulses led to maximum increases of α-carotene, 9- and 13-cis-lycopene, which increased by 93%, 94% and 140%, respectively. Therefore, MIPEF could stimulate synthesis of secondary metabolites and contribute to production of tomatoes with high individual polyphenol and carotenoid contents.

Effects of Pulsed Electric Fields on the Bioactive Compound Content and Antioxidant Capacity of Tomato Fruit

The effect of moderate intensity pulsed electric fields (MIPEF) on the bioactive compounds (total polyphenol, lycopene, and vitamin C content) as well as on the antioxidant capacity of tomato fruit was studied. The MIPEF treatment conditions were optimized to obtain tomato fruit with a high content of bioactive compounds. Tomato fruits were subjected to different electric field strengths (from 0.4 to 2.0 kV/cm) and number of pulses (from 5 to 30) and then immediately refrigerated at 4 °C for 24 h. A concentration of bioactive compounds higher than that of untreated tomatoes was obtained in MIPEF-treated tomatoes. A 44% increase in total polyphenol content was achieved under 30 pulses at 1.2 kV/cm. The hydrophilic antioxidant capacity was also enhanced by 44% applying 18 pulses at 1.2 kV/cm, and the lipophilic antioxidant capacity was increased by 37% under 5 pulses at 1.2 kV/cm. The maximum overall level of bioactive compounds and antioxidant capacity in the treated tomatoes was obtained under 16 pulses at 1 kV/cm. Therefore, MIPEF treatments could be considered an effective method to enhance the bioactive compound content and antioxidant potential of tomatoes.

A metabolomic approach differentiates between conventional and organic ketchups.

The agronomic environments in which tomatoes are cultivated potentially affect the levels of antioxidants and other metabolites in commercial products. In this study, biochemical and metabolomic techniques were used to assess the differences between ketchups produced by organic and conventional systems. An untargeted metabolomic approach using QToF-MS was used to identify those nutrients that have the greatest impact on the overall metabolomic profile of organic ketchups as compared to conventional ones. Individual polyphenols were quantified using LC-ESI-QqQ. This multifaceted approach revealed that the agronomic environment in which tomatoes are grown induces alterations in the content of antioxidant capacity, phenolics, and other metabolites in ketchups. Organic cultivation was found to provide tomatoes and tomato-derived products with a significantly higher content of antioxidant microconstituents, whereas glutamylphenylalanine and N-malonyltryptophan were detected only in conventional ketchups.

Impact of high-intensity pulsed electric fields on carotenoids profile of tomato juice made of moderate-intensity pulsed electric field-treated tomatoes.

The effect of pulsed electric fields (PEF) on the carotenoid content of tomato juices was studied. First, moderate-intensity PEF (MIPEF) was applied to raw tomatoes. Afterwards, MIPEF-treated and untreated tomatoes were immediately refrigerated at 4 °C for 24 h and then, they were separately ground to produce tomato juices. Juices were treated by heat treatments or by high-intensity PEF (HIPEF) and stored under refrigeration for 56 days. MIPEF treatment of tomatoes increased the content of carotenoid compounds in tomato juices. An enhancement of 63-65% in 15-cis-lycopene was observed in juices prepared with MIPEF-treated tomatoes. A slight increase in cis-lycopene isomers was observed over time, whereas other carotenoids slightly decreased. However, HIPEF treated tomato juices maintained higher carotenoid content (10-20%) through the storage time than thermally and untreated juices. The combination of MIPEF and HIPEF treatments could be used not only to produce tomato juices with high carotenoid content but also, to maintain higher the carotenoid content during storage time.

Effects of alcohol and polyphenols from beer on atherosclerotic biomarkers in high cardiovascular risk men: A randomized feeding trial

BACKGROUND AND AIMS Moderate alcohol consumption exerts a cardioprotective effect, but no studies have evaluated the alcohol-independent cardiovascular effects of the non-alcoholic components of beer. We aimed to evaluate the effects of ethanol and the phenolic compounds of beer on classical and novel cardiovascular risk factors. METHODS AND RESULTS Thirty-three high risk male volunteers were included in a randomized, crossover feeding trial. After a washout period, all subjects received beer (30 g alcohol/d, 660 mL), the equivalent amount of polyphenols as non-alcoholic beer (990 mL), and gin (30 g alcohol/d, 100 mL) for 4 weeks. All outcomes were evaluated before and after each intervention period. Moderate alcohol consumption increased serum HDL-cholesterol (∼5%), ApoA-I (∼6%), ApoA-II (∼7%) and adiponectin (∼7%), and decreased serum fibrinogen (∼8%), and interleukin (IL)-5 (∼14%) concentrations, whereas the non-alcoholic fraction of beer (mainly polyphenols) increased the receptor antagonist of IL-1 (∼24%), and decreased lymphocyte expression of lymphocyte function-associated antigen-1 (∼11%), lymphocyte and monocyte expression of Sialil-Lewis X (∼16%) and monocyte expression of CCR2 (∼31%), and tumor necrosis factor (TNF)-β (∼14%) and IL-15 (∼22%) plasma concentrations. No changes were observed in glucose metabolism parameters or in body weight and adiposity parameters. CONCLUSION The phenolic content of beer reduces leukocyte adhesion molecules and inflammatory biomarkers, whereas alcohol mainly improves the lipid profile and reduces some plasma inflammatory biomarkers related to atherosclerosis.