Metabolic Profiles and Blood Biomarkers to Discriminate between Benign Thyroid Nodules and Papillary Carcinoma, Based on UHPLC-QTOF-ESI+-MS Analysis.

In this study, serum metabolic profiling of patients diagnosed with papillary thyroid carcinoma (PTC) and benign thyroid pathologies (BT) aimed to identify specific biomarkers and altered pathways when compared with healthy controls (C). The blood was collected after a histological confirmation from PTC (n = 24) and BT patients (n = 31) in parallel with healthy controls (n = 81). The untargeted metabolomics protocol was applied by UHPLC-QTOF-ESI+-MS analysis and the statistical analysis was performed using the MetaboAnalyst 5.0 platform. The partial least squares-discrimination analysis, including VIP values, random forest graphs, and heatmaps (p < 0.05), was complemented with biomarker analysis (with AUROC ranking) and pathway analysis, suggesting a model for abnormal metabolic pathways in PTC and BT based on 166 identified metabolites. There were 11 classes of putative biomarkers selected that were involved in altered metabolic pathways, e.g., polar molecules (amino acids and glycolysis metabolites, purines and pyrimidines, and selenium complexes) and lipids including free fatty acids, bile acids, acylated carnitines, corticosteroids, prostaglandins, and phospholipids. Specific biomarkers of discrimination were identified in each class of metabolites and upregulated or downregulated comparative to controls, PTC group, and BT group. The lipidomic window was revealed to be more relevant for finding biomarkers related to thyroid carcinoma or benign thyroid nodules, since our study reflected a stronger involvement of lipids and selenium-related molecules in metabolic discrimination.

Potential Diagnostic Biomarker Detection for Prostate Cancer Using Untargeted and Targeted Metabolomic Profiling.

Prostate cancer (PCa) remains one of the leading causes of cancer mortality in men worldwide, currently lacking specific, early detection and staging biomarkers. In this regard, modern research focuses efforts on the discovery of novel molecules that could represent potential future non-invasive biomarkers for the diagnosis of PCa, as well as therapeutic targets. Mounting evidence shows that cancer cells express an altered metabolism in their early stages, making metabolomics a promising tool for the discovery of altered pathways and potential biomarker molecules. In this study, we first performed untargeted metabolomic profiling on 48 PCa plasma samples and 23 healthy controls using ultra-high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-[ESI+]-MS) for the discovery of metabolites with altered profiles. Secondly, we selected five molecules (L-proline, L-tryptophan, acetylcarnitine, lysophosphatidylcholine C18:2 and spermine) for the downstream targeted metabolomics and found out that all the molecules, regardless of the PCa stage, were decreased in the PCa plasma samples when compared to the controls, making them potential biomarkers for PCa detection. Moreover, spermine, acetylcarnitine and L-tryptophan had very high diagnostic accuracy, with AUC values of 0.992, 0.923 and 0.981, respectively. Consistent with other literature findings, these altered metabolites could represent future specific and non-invasive candidate biomarkers for PCa detection, which opens novel horizons in the field of metabolomics.

Enhanced diagnosis and prognosis of severe alcoholic hepatitis using novel metabolomic biomarkers.

AIM: Differentiating alcoholic hepatitis (AH) from acute decompensation of alcoholic cirrhosis (DC) is challenging, as the presentation and biochemistry are similar. We aimed to identify potential metabolomic biomarkers to differentiate between AH and DC, and to predict short-term mortality. METHODS: We included consecutive biopsy proven AH and DC patients, which were managed according to current guidelines and followed up until the end of the study. Untargeted metabolomics was assessed in all patients at baseline. Specific analyses were successively performed to identify potential biomarkers, which were further semi-quantitatively analysed against relevant clinical endpoints. RESULTS: Thirty-four patients with AH and 37 with DC were included. UHPLC-MS analysis identified 83 molecules potentially differentiating between AH and DC. C16-Sphinganine-1P (S1P) was the most increased, whereas Prostaglandin E2 (PGE2) was the most decreased. The PGE2/S1P ratio < 1.03 excellently discriminates between AH and DC: AUC 0.965 (p < 0.001), Se 90%, Sp 100%, PPV 0.91, NPV 1, and diagnostic accuracy 95%. This ratio is not influenced by the presence of infection (AUC 0.967 vs. 0.962), correlates with the Lille score at 7 days (r = -0.60; P = 0.022) and tends to be lower in corticosteroid non-responders as compared with patients who responded [0.85(±0.02) vs. 0.89(±0.05), P = 0.069]. Additionally, decreased ursodeoxycholic acid levels are correlated with MELD and Maddrey scores and predict mortality with a 77.27% accuracy (NPV = 100%). CONCLUSION: This study suggests the PGE2 (decreased)/S1P (increased) ratio as a biomarker to differentiate AH from DC. The study also finds that low levels of ursodeoxycholic acid could predict increased mortality in AH.

Pentacyclic Triterpenoid Phytochemicals with Anticancer Activity: Updated Studies on Mechanisms and Targeted Delivery.

Pentacyclic triterpenoids (TTs) represent a unique family of phytochemicals with interesting properties and pharmacological effects, with some representatives, such as betulinic acid (BA) and betulin (B), being mainly investigated as potential anticancer molecules. Considering the recent scientific and preclinical investigations, a review of their anticancer mechanisms, structure-related activity, and efficiency improved by their insertion in nanolipid vehicles for targeted delivery is presented. A systematic literature study about their effects on tumor cells in vitro and in vivo, as free molecules or encapsulated in liposomes or nanolipids, is discussed. A special approach is given to liposome-TTs and nanolipid-TTs complexes to be linked to microbubbles, known as contrast agents in ultrasonography. The production of such supramolecular conjugates to deliver the drugs to target cells via sonoporation represents a new scientific and applicative direction to improve TT efficiency, considering that they have limited availability as lipophilic molecules. Relevant and recent examples of in vitro and in vivo studies, as well as the challenges for the next steps towards the application of these complex delivery systems to tumor cells, are discussed, as are the challenges for the next steps towards the application of targeted delivery to tumor cells, opening new directions for innovative nanotechnological solutions.

Metabolite Profiling of the Gut-Renal-Cerebral Axis Reveals a Particular Pattern in Early Diabetic Kidney Disease in T2DM Patients.

Type 2 diabetes mellitus (T2DM) represents an important microvascular disease concerning the kidney and the brain. Gut dysbiosis and microbiota-derived metabolites may be in relation with early pathophysiological changes in diabetic kidney disease (DKD). The aim of the study was to find new potential gut-derived biomarkers involved in the pathogenesis of early DKD, with a focus on the complex interconnection of these biomarkers with podocyte injury, proximal tubule dysfunction, renal and cerebrovascular endothelial dysfunction. The study design consisted of metabolite profiling of serum and urine of 90 T2DM patients (subgroups P1-normoalbuminuria, P2-microalbuminuria, P3-macroalbuminuria) and 20 healthy controls (group C), based on ultra-high-performance liquid chromatography coupled with electrospray ionization-quadrupole-time of flight-mass spectrometry analysis (UHPLC-QTOF-ESI+-MS). By multivariate and univariate analyses of serum and urine, which included Partial Least Squares Discriminant Analysis (PLSDA), Variable Importance Plots (VIP), Random Forest scores, One Way ANOVA and Biomarker analysis, there were discovered metabolites belonging to nitrogen metabolic pathway and retinoic acid signaling pathway which differentiate P1 group from P2, P3, C groups. Tyrosine, phenylalanine, indoxyl sulfate, serotonin sulfate, and all-trans retinoic acid express the metabolic fingerprint of P1 group vs. P2, P3, C groups, revealing a particular pattern in early DKD in T2DM patients.

Stabilization of Sunflower Oil with Biologically Active Compounds from Berries.

Sunflower oil (Helianthus annuus) contains a rich concentration of polyunsaturated fatty acids, which are susceptible to rapid oxidative processes. The aim of this study was to evaluate the stabilizing effect of lipophilic extracts from two types of berries, sea buckthorn and rose hips, on sunflower oil. This research included the analysis of sunflower oil oxidation products and mechanisms, including the determination of chemical changes occurring in the lipid oxidation process via LC-MS/MS using electrospray ionization in negative and positive mode. Pentanal, hexanal, heptanal, octanal, and nonanal were identified as key compounds formed during oxidation. The individual profiles of the carotenoids from sea buckthorn berries were determined using RP-HPLC. The influence of the carotenoid extraction parameters ascertained from the berries on the oxidative stability of sunflower oil was analyzed. The dynamics of the accumulation of the primary and secondary products of lipid oxidation and the variation of the carotenoid pigment content in the lipophilic extracts of sea buckthorn and rose hips during storage demonstrated good stability at 4 °C in the absence of light for 12 months. The experimental results were applied to mathematical modeling using fuzzy sets and mutual information analysis, which allowed for the prediction of the oxidation of sunflower oil.

Anthocyanins as Key Phytochemicals Acting for the Prevention of Metabolic Diseases: An Overview.

Anthocyanins are water-soluble pigments present in fruits and vegetables, which render them an extensive range of colors. They have a wide distribution in the human diet, are innocuous, and, based on numerous studies, have supposed preventive and therapeutical benefits against chronic affections such as inflammatory, neurological, cardiovascular, digestive disorders, diabetes, and cancer, mostly due to their antioxidant action. Despite their great potential as pharmaceutical applications, they have a rather limited use because of their rather low stability to environmental variations. Their absorption was noticed to occur best in the stomach and small intestine, but the pH fluctuation of the digestive system impacts their rapid degradation. Urine excretion and tissue distribution also occur at low rates. The aim of this review is to highlight the chemical characteristics of anthocyanins and emphasize their weaknesses regarding bioavailability. It also targets to deliver an update on the recent advances in the involvement of anthocyanins in different pathologies with a focus on in vivo, in vitro, animal, and human clinical trials.

Extracts of the Wild Potato Species Solanum chacoense on Breast Cancer Cells: Biochemical Characterization, In Vitro Selective Cytotoxicity and Molecular Effects.

Solanum chacoense (wild potato) is intensively used in breeding, its biochemical profile and putative human health-related traits being transferred into potato cultivars aimed for consumption. The goal of this study was to evaluate the biochemical profile and the anti-tumor potential of methanolic extracts obtained from S. chacoense leaves and tubers against three breast cancer cell lines in comparison to healthy endothelial cells (HUVEC). The biochemical profile of the extracts was determined by HPLC-PDA/-ESI+-MS and ITEX/GC-MS, the selective cytotoxicity by MTT assay whereas RT-qPCR was used to evaluate the expression of proliferation- and apoptosis-related genes. Both extracts proved to be rich in phenolic acids and volatile compounds, the leaf extract also containing glycoalkaloids. Both extracts proved to be cytotoxic for breast cancer cell lines, with IC50 values varying between 132.9 and 390.7 µg/ml. Both extracts had selective cytotoxicity against MCF7 cell line in comparison to HUVECs (selectivity coefficients >2.3). The treatment with the extracts induced overexpression of the pro-apoptotic gene BAX¸ down-regulation of the anti-apoptotic gene BCL-2 and the pro-proliferation genes NFkB, CCND1, and STAT3. Thus S. chacoense extracts proved to be rich in compounds with anticancer proprieties and are capable of inducing selective cytotoxicity on MCF7 cell line.

ATR-FTIR-MIR Spectrometry and Pattern Recognition of Bioactive Volatiles in Oily versus Microencapsulated Food Supplements: Authenticity, Quality, and Stability.

Fourier transform infrared spectroscopy on the middle infrared region (ATR-FTIR-MIR) proved to be a convenient and reliable technique to evaluate foods' quality and authenticity. Plants' essential oils are bioactive mixtures used as such or in different oily or microencapsulated formulations, beneficial to human health. Six essential oils (thyme, oregano, juniperus, tea tree, clove, and cinnamon) were introduced in three oily formulations (Biomicin, Biomicin Forte, and Biomicin urinary) and these formulations were microencapsulated on fructose and maltodextrin matrices. To study their stability, the microencapsulated powders were kept under light irradiation for 14 days at 25 °C or introduced in biopolymer capsules. All variants were analysed by ATR-FTIR-MIR, recording wavenumbers and peak intensities (3600-650 cm-1). The data were processed by Unscrambler and Metaboanalyst software, with specific algorithms (PCA, PLSDA, heatmaps, and random forest analysis). The results demonstrated that ATR-FTIR-MIR can be successfully applied for fingerprinting and finding essential oil biomarkers as well as to recognize this pattern in final microencapsulated food supplements. This study offers an improved ATR-FTIR-MIR procedure coupled with an adequate chemometric analysis and accurate data interpretation, to be applied for the evaluation of authenticity, quality, traceability, and stability during storage of essential oils incorporated in different matrices.

The Influence of Temperature, Storage Conditions, pH, and Ionic Strength on the Antioxidant Activity and Color Parameters of Rowan Berry Extracts.

Recent trends in the food industry combined with novel methods in agriculture could transform rowan into a valuable raw material with potential technological applications. Thus, the aim of this research was to investigate the content of bioactive compounds in its fruits and to assess the color and antioxidant stability of the extracts prepared from such fruits during various thermal treatments and at different pH and ionic strength values. Various spectrophotometric methods, HPLC, and capillary electrophoresis were used to quantify the concentrations of bioactive compounds-polyphenols, carotenoids, organic acids, and to assess antioxidant activity and color. The results show that rowan berries contain circa 1.34-1.47 g/100 g of polyphenols among which include catechin, epicatechin, ferulic acid methyl ester, procyanidin B1, etc.; ca 21.65 mg/100 g of carotenoids including zeaxanthin, ß-cryptoxanthin, all-trans-ß-carotene, and various organic acids such as malic, citric, and succinic, which result in a high antioxidant activity of 5.8 mmol TE/100 g. Results also showed that antioxidant activity exhibited high stability when the extract was subjected to various thermal treatments, pHs, and ionic strengths, while color was mainly impacted negatively when a temperature of 100 °C was employed. This data confirms the technological potential of this traditional, yet often overlooked species.

Comparison of Different Extraction Solvents for Characterization of Antioxidant Potential and Polyphenolic Composition in Boletus edulis and Cantharellus cibarius Mushrooms from Romania.

Edible mushrooms are well-known for their nutritional benefits and low energy density. In addition, mushroom extracts contain various bioactive compounds that account for their antioxidant activity; the applied extraction conditions influence the extraction efficiency of such compounds. Therefore, this study investigates the effects of four extractants on the content of polyphenols and antioxidant properties of Boletus edulis and Cantharellus cibarius mushrooms, aiming to optimize the extraction process. Powders of B. edulis and C. cibarius mushrooms were subjected to extraction with acidic water (10% CH3COOH), ethanol/water/acetic acid (15:76.5:8.5, v/v/v), hexane, and diethyl ether to measure their total phenolic content (TPC), total flavonoid content (TFC), and Trolox equivalent antioxidant capacity (TEAC). Furthermore, the level of individual polyphenolic compounds in these extracts was quantified using an HPLC-DAD-ESI-MS method. Results showed that the type of solvent significantly influenced the TPC and TEAC of mushroom powder but insignificantly influenced the TFC. A very strong positive correlation was found between TPC and TEAC, but no correlation was found between TFC and TEAC or TPC and TFC. Acidic water extracted the highest amount of polyphenolic compounds from these mushroom powders. Therefore, the aqueous extract showed the highest TPC and strongest antioxidant activity. Thus, acidic water is recommended for polyphenol analysis in B. edulis and C. cibarius mushrooms.

Melatonin, an ubiquitous metabolic regulator: functions, mechanisms and effects on circadian disruption and degenerative diseases.

The last four decades, we assist to an increasing scientific interest on melatonin, a circadian hormone, a metabolic regulator which influences not only plants' metabolism and their defense against pathogens but mostly the animals and humans' metabolic pathways, their response to circadian disruption, stress and burnout syndrome. In humans, as a hormonal regulator, produced in the pineal grand as well in mitochondria, melatonin is involved in different, complex intracellular signaling pathways, with antioxidant and immune stimulating effects, proving to act as a circadian synchronizer, as a preventive and therapeutic agent in many degenerative diseases, and especially in hormone-dependent cancers. Preclinical or clinical studies showed recently the mechanisms involved in regulating the cellular activity, its role in aging and circadian disturbances and impact on degenerative diseases. Melatonin proved to have an anti-inflammatory, antiapoptotic and powerful antioxidant effect by subtle mechanisms in mitochondrial metabolic pathways. This overview includes recent and relevant literature data related to the impact of endogenous and exogeneous melatonin on the prevention of cancer progression and treatment of various degenerative diseases. Metabolomics, an emerging new omics' technology, based on high performance liquid chromatography coupled with mass spectrometry is presented as an encouraging technique to fingerprint and realize a precise evaluation and monitoring of the turnover of melatonin and its metabolites in different pathological circumstances.

The decrease of some serum free amino acids can predict breast cancer diagnosis and progression.

This study was targeted on a metabolomic approach to compare the blood serum free amino acid profiles and concentration of confirmed breast cancer (stages I-III) patients to healthy controls in order to establish reliable biomarkers of early detection and prediction of breast cancer. The ultra-high-performance liquid chromatography coupled with mass spectrometry using positive ionization electrospray was applied for the picoline-derivatized serum free amino acids using the EZ:faastTM kit. Multivariate statistical analysis principal component analysis, partial least squares discrimination analysis and univariate analysis were applied in order to discriminate between patient groups and putative amino acid biomarkers for breast cancer. A significant decrease of amino acid concentrations between the breast cancer group and the control group was positively correlated with breast cancer progression. Arginine, Alanine, Isoleucine, Tyrosine and Tryptophan were identified as being good potential discriminants (AUROC ≥0.85) and suitable candidates to diagnose and predict the breast cancer progression.

Lipidomics biomarkers in women with polycystic ovary syndrome (PCOS) using ultra-high performance liquid chromatography-quadrupole time of flight electrospray in a positive ionization mode mass spectrometry.

Polycystic ovary syndrome (PCOS), characterized by oligo-anovulation and androgen excess is considered a high-risk condition for metabolic disorders. Herein, untargeted metabolomics analysis was applied to women with PCOS, aiming to provide deeper insights into lipidomics biomarkers signature of PCOS, for better diagnosis and management. This was a cross-sectional study in which 15 Caucasian women with PCOS and 15 Caucasian healthy, age-matched women were enrolled. Lipidomics analysis was performed using Ultra-High Performance Liquid Chromatography-Quadrupole Time of Flight Electrospray Mass Spectrometry. Partial Least Squares Discriminant Analysis retrieved the most important discriminative metabolites. Significantly increased levels of triacylglycerol (18:2/18:2/0-18:0) in addition to cholestane-3beta, 5alpha, 6beta-triol (18:0/0:0) and cholestane-5alpha (18:1/0:0) appeared as valuable variables to differentiate subjects with PCOS from controls. Acyl-carnitine 2-hydroxylauroylcarnitine was significantly elevated in PCOS in opposition to decreased phosphocholines metabolites (18:1/18:4, 18:3/18:2), to suggest a metabolic pattern linked to lipid peroxidation. A high fat intake or reduced fat energy consumption during nighttime due to diminished ability to switch to lipid oxidation during fasting time possibly contribute to hypertriglyceridemia found in PCOS. Furthermore, inflammatory mediators including metabolites of the prostaglandin (PG) E2 pathway and oxo-leukotrienes (LT) were increased in patients with PCOS. Potential lipidomics biomarkers were identified that could stratify between women with PCOS and healthy controls. The results show particular alterations in acylglycerols, PGs and LTs and phosphocholines and carnitine metabolites. The lipidomics profiles of PCOS indicate a higher risk of developing metabolic diseases.

Serum metabolomics can predict the outcome of first systematic transrectal prostate biopsy in patients with PSA <10 ng/ml.

AIM: To assess the predictive value of metabolomic analysis for the presence of prostate cancer (PCa) at first systematic biopsy. PATIENTS & METHODS: Ninety serum samples from patients with suspicion for PCa were included. Targeted and nontargeted metabolomic analysis was performed. RESULTS: Six metabolites were combined into a predictive score. A cutoff value of 0.528 for the metabolomic score showed a good accuracy for the prediction of PCa at biopsy (Area under the curve (AUC): 0.779; p < 0.001). These results were validated in a subgroup of patients, showing similar accuracy (p = 0.1). For patients with prostate specific antigen (PSA) less than 10 ng/ml, the score showed a Se 80.95%, Sp 64.52% for the detection of PCa at biopsy. CONCLUSION: Metabolomic analysis can predict the outcome of the first systematic biopsy.

Characterization of a Sea Buckthorn Extract and Its Effect on Free and Encapsulated Lactobacillus casei.

Probiotics are bacteria that can provide health benefits to consumers and are suitable to be added to a variety of foods. In this research, viability of immobilized Lactobacillus casei in alginate with or without sea buckthorn lipid extract were studied during heat treatment and with an in vitro gastrointestinal model. The characterization of the lipid extract was also done using the UV-Vis spectrometry (UV-Vis), high-performance liquid chromatography photodiode array detection method (HPLC-PDA), gas chromatography coupled with mass spectrometry (GS-MS) and Cryo scanning electron microscopy (Cryo-SEM). During heat treatment, the entrapped probiotic cells proved high viability (>6 CFU log/g), even at temperatures above 50 °C. The rich in monounsaturated fatty acids sea buckthorn fraction improved the in vitro digestion passage regarding the probiotic viability. The survival of the probiotic cells was 15% higher after 2 h in the acidic medium of the simulated gastric fluid in the sample where L. casei was encapsulated with the sea buckthorn extract compared with the samples where no extract was added. Thus, this approach may be effective for the future development of probiotic-supplemented foods as foods with health welfare for the consumers.

Melanoma Inhibition by Anthocyanins Is Associated with the Reduction of Oxidative Stress Biomarkers and Changes in Mitochondrial Membrane Potential.

Anthocyanins are water soluble pigments which have been proved to exhibit health benefits. Several studies have investigated their effects on several types of cancer, but little attention has been given to melanoma. The phytochemical content of nine different berry samples was assessed by liquid chromatography followed by electrospray ionization mass spectrometry (LC-ESI+-MS). Twenty-six anthocyanins were identified, after a previous C18 Sep-pak clean-up procedure. Chokeberry and red grape anthocyanins rich extracts (C-ARE and RG-ARE) were selected to be tested on normal and melanoma cell lines, due to their different chemical pattern. C-ARE composition consists of cyanidin aglycone glycosylated with different sugars; while RG-ARE contains glucosylated derivatives of five different aglycones. Both C-ARE and RG-ARE anthocyanins reduced proliferation, increased oxidative stress biomarkers and diminished mitochondrial membrane potential in melanoma cells, having no negative influence on normal cells. A synergistic response may be attributed to the five different aglycones present in RG-ARE, which proved to exert greater effects on melanoma cells than the mixture of cyanidin derivatives with different sugars (C-ARE). In conclusion, C-ARE and RG-ARE anthocyanins may inhibit melanoma cell proliferation and increase the level of oxidative stress, with opposite effect on normal cells. Therefore, anthocyanins might be recommended as active ingredients for cosmetic and nutraceutical industry. Graphical Abstract ᅟ.

Antiproliferative and antioxidant properties of anthocyanin rich extracts from blueberry and blackcurrant juice.

The present study was aimed at evaluating the antiproliferative potential of anthocyanin-rich fractions (ARFs) obtained from two commercially available juices (blueberry and blackcurrant juices) on three tumor cell lines; B16F10 (murine melanoma), A2780 (ovarian cancer) and HeLa (cervical cancer). Individual anthocyanin determination, identification and quantification were done using HPLC-MS. Antioxidant activity of the juices was determined through different mechanism methods such as DPPH and ORAC. For biological testing, the juices were purified through C18 cartridges in order to obtain fractions rich in anthocyanins. The major anthocyanins identified were glycosylated cyanidin derivatives. The antiproliferative activity of the fractions was tested using the MTT assay. The antiproliferative potential of ARF was found to be associated with those bioactive molecules, anthocyanins due to their antioxidant potential. The results obtained indicated that both blueberry and blackcurrants are rich sources of antioxidants including anthocyanins and therefore these fruits are highly recommended for daily consumption to prevent numerous degenerative diseases.

The influence of different polymers on viability of Bifidobacterium lactis 300b during encapsulation, freeze-drying and storage.

Seven different types of natural polymers namely hydroxypropyl methylcellulose (HPMC), sodium-carboxymethyl cellulose (Na-CMC), microcrystalline cellulose (MCC), starch BR-07, starch BR-08, dextrin and pullulan were used in order to develop the optimal formula for the entrapment of Bifidobacterium lactis 300B in Ca-alginate based granules. Laminar flow drip casting with Brace-Encapsulator was used in order to prepare the granules. The results showed that alginate/pullulan and alginate/HPMC formulation provide high protection for the bacterial strain used for encapsulation. These two formulations were further used to obtain freeze dried granules, for which the viability in time and at different temperatures was tested. The final results showed a higher viability than the level of the therapeutic minimum (>10(7) CFU/g) after 15 days of storage. Other parameters like entrapment efficiency, production rate, sphericity, flowability were also discussed.

Chemometric discrimination of different tomato cultivars based on their volatile fingerprint in relation to lycopene and total phenolics content.

INTRODUCTION: The characteristic flavour of tomato is given by a complex mixture of sugars, acids, amino acids, minerals and volatile metabolites. Of these, volatile compounds are considered to greatly influence the flavour of tomato fruits. The volatile aroma compounds and phytochemical content of tomatoes are dependent on genotype, environmental conditions and cultural practices, and can thus be used for cultivar discrimination. OBJECTIVE: To assess the possibility of using the volatile profile of tomato to fingerprint and discriminate different tomato cultivars based on an 'in-tube extraction' technique coupled to gas chromatography, combined with mass spectrometry (ITEX/GC-MS) and a chemometric approach. RESULTS: Using the ITEX/GC-MS technique, 61 volatiles were analysed and separated from tomato cultivars, with 58 being identified. The main volatiles identified in all tomato cultivars were: hexanal, trans-2-hexenal, 1-hexanol, 3-pentanone, 3-methylbutanol, 2-methylbutanol, 3-methylbutanal and 6-methyl-5-hepten-2-one. The lycopene content and total phenolic compound content of the tomato cultivars varied between 36.78 and 73.18 mg/kg fresh weight (fw) and from 119.4 to 253.7 mg of gallic acid equivalents (GAE) per kilogram fresh weight, respectively. Volatile fingerprint and phytochemical composition led to a good differentiation between tomato cultivars, with the first two principal components explaining 89% of the variance in the data. CONCLUSION: The tomato cultivars studied were easily discriminated based on their characteristic volatile profile that was obtained using the reliable ITEX/GC-MS technique. Principal component analysis revealed, in addition to volatile compounds, the important role played by the total phenolic content in tomato cultivar discrimination, which is highly correlated with phenotypic and biochemical differences between tomato cultivars.