Corrigendum to "Phytochemicals and bioactive constituents in food packaging - A systematic review" [Heliyon 9(11) (November 2023) e21196].

[This corrects the article DOI: 10.1016/j.heliyon.2023.e21196.].

Phytochemicals and bioactive constituents in food packaging - A systematic review.

Designing and manufacturing functional bioactive ingredients and pharmaceuticals have grown worldwide. Consumers demand for safe ingredients and concerns over harmful synthetic additives have prompted food manufacturers to seek safer and sustainable alternative solutions. In recent years the preference by consumers to natural bioactive agents over synthetic compounds increased exponentially, and consequently, naturally derived phytochemicals and bioactive compounds, with antimicrobial and antioxidant properties, becoming essential in food packaging field. In response to societal needs, packaging needs to be developed based on sustainable manufacturing practices, marketing strategies, consumer behaviour, environmental concerns, and the emergence of new technologies, particularly bio- and nanotechnology. This critical systematic review assessed the role of antioxidant and antimicrobial compounds from natural resources in food packaging and consumer behaviour patterns in relation to phytochemical and biologically active substances used in the development of food packaging. The use of phytochemicals and bioactive compounds inside packaging materials used in food industry could generate unpleasant odours derived from the diffusion of the most volatile compounds from the packaging material to the food and food environment. These consumer concerns must be addressed to understand minimum concentrations that will not affect consumer sensory and aroma negative perceptions. The research articles were carefully chosen and selected by following the Preferred Reporting Items for Systematic Reviews (PRISMA) guidelines.

Tracing the volatilomic fingerprint of grape pomace as a powerful approach for its valorization.

The huge amount of grape pomace (GP) generated every year worldwide, particularly in Europe, creates negative impacts at the economic and environmental levels. As far as we know, scarce research has been done on the volatilomic fingerprint of GP. To meet consumer demand for healthy foods, there is a growing interest in the characterization of particular volatile organic metabolites (VOMS) in GP that can be used for industrial applications, including the food industry. In this study, the volatilomic fingerprint of GP obtained from different Vitis vinifera L. grapes was established by solid phase microextraction (HS-SPME) combined to gas chromatography-mass spectrometry (GC-MS), to explore the properties of most dominant VOMs in a context of its application on marketable products. A total of 52 VOMs belonging to different chemical families were identified. Alcohols, carbonyl compounds, and esters, are the most dominant, representing 38.8, 29.3, and 24.2% of the total volatile profile of the investigated GP, respectively. Esters (e.g., isoamyl acetate, hexyl acetate, ethyl hexanoate) and alcohols (e.g., 3-methyl butan-2-ol, hexan-1-ol) can be used as flavoring agents with potential use in the food industry, and in the cosmetic industry, for fragrances production. In addition, the identified terpenoids (e.g., menthol, ylangene, limonene) exhibit antioxidant, antimicrobial, and anticancer, biological properties, among others, boosting their potential application in the pharmaceutical industry. The obtained results revealed the potential of some VOMs from GP to replace synthetic antioxidants, colorants, and antimicrobials used in the food industry, and in the cosmetic and pharmaceutical industry, meeting the increasing consumer demand for natural alternative compounds.

The Fingerprint of Fortified Wines-From the Sui Generis Production Processes to the Distinctive Aroma.

The fortified wines that originated in Mediterranean countries have, in common, a high alcohol content to increase their shelf-life during long journeys to northern Europe and the American continent. Nowadays, the world's better-known wines, including Marsala, Madeira, Port, and Sherry, due to their high alcoholic content, sweet taste, and intense aromatic profile, are designated as dessert wines and sometimes served as aperitifs. This review gives an overview of the traditional vinification process, including the microbiota and autochthonous yeast, as well as the regulatory aspects of the main Italian, Portuguese, and Spanish fortified wines. The winemaking process is essential to defining the volatile organic compounds (VOCs) that characterize the aroma of each fortified wine, giving them an organoleptic fingerprint and "terroir" characteristics. The various volatile and odorous compounds found in fortified wines during the oxidative aging are discussed in the last part of this review.

Differences in the Volatilomic Urinary Biosignature of Prostate Cancer Patients as a Feasibility Study for the Detection of Potential Biomarkers.

Prostate cancer (PCa) continues to be the second most common malignant tumour and the main cause of oncological death in men. Investigating endogenous volatile organic metabolites (VOMs) produced by various metabolic pathways is emerging as a novel, effective, and non-invasive source of information to establish the volatilomic biosignature of PCa. In this study, headspace solid-phase microextraction combined with gas chromatography-mass spectrometry (HS-SPME/GC-MS) was used to establish the urine volatilomic profile of PCa and identify VOMs that can discriminate between the two investigated groups. This non-invasive approach was applied to oncological patients (PCa group, n = 26) and cancer-free individuals (control group, n = 30), retrieving a total of 147 VOMs from various chemical families. This included terpenes, norisoprenoid, sesquiterpenes, phenolic, sulphur and furanic compounds, ketones, alcohols, esters, aldehydes, carboxylic acid, benzene and naphthalene derivatives, hydrocarbons, and heterocyclic hydrocarbons. The data matrix was subjected to multivariate analysis, namely partial least-squares discriminant analysis (PLS-DA). Accordingly, this analysis showed that the group under study presented different volatomic profiles and suggested potential PCa biomarkers. Nevertheless, a larger cohort of samples is required to boost the predictability and accuracy of the statistical models developed.

Tracing the Volatilomic Fingerprint of the Most Popular Italian Fortified Wines.

The aim of the current study was to provide a useful platform to identify characteristic molecular markers related to the authenticity of Italian fortified wines. For this purpose, the volatilomic fingerprint of the most popular Italian fortified wines was established using headspace solid-phase microextraction combined with gas chromatography-mass spectrometry (HS-SPME/GC-MS). Several volatile organic compounds (VOCs), belonging with distinct chemical groups, were identified, ten of which are common to all the analyzed fortified Italian wines. Terpenoids were the most abundant chemical group in Campari bitter wines due to limonene's high contribution to the total volatilomic fingerprint, whereas for Marsala wines, alcohols and esters were the most predominant chemical groups. The fortified Italian wines VOCs network demonstrated that the furanic compounds 2-furfural, ethyl furoate, and 5-methyl-2-furfural, constitute potential molecular markers of Marsala wines, while the terpenoids nerol, α-terpeniol, limonene, and menthone isomers, are characteristic of Vermouth wines. In addition, butanediol was detected only in Barolo wines, and ß-phellandrene and ß-myrcene only in Campari wines. The obtained data reveal an adequate tool to establish the authenticity and genuineness of Italian fortified wines, and at the same time constitute a valuable contribution to identify potential cases of fraud or adulteration to which they are subject, due to the high commercial value associated with these wines. In addition, they contribute to the deepening of scientific knowledge that supports its valorization and guarantee of quality and safety for consumers.

Impact of Japanese beetles (Popillia japonica Newman) on the chemical composition of two grape varieties (Nebbiolo and Erbaluce) grown in Italy.

The effect of Japanese beetles (P. japonica) on critical quality indicators of Nebbiolo and Erbaluce grapes, specifically on their phenolic and volatile composition, was assessed. Adult beetle symptoms include extended skeletonization of leaves. Leaves are frequently left with their mid-vein intact but, when severely damaged, quickly turn brown. However, the plant tends to recover by generating a new leaf apparatus and the grapes reach ripeness. It emerged that the phenolic content of grapes produced by plants attacked by P. japonica (396 and 550 mg/kg, Nebbiolo and Erbaluce respectively) was generally higher when compared to healthy plants (266 and 188 mg/kg, Nebbiolo and Erbaluce respectively). Similarly, in the (red) Nebbiolo cultivar, the anthocyanin content was significantly lower in grapes produced with healthy plants. The influence of P. japonica on the volatile composition of Nebbiolo and Erbaluce grapes showed a total volatile fraction of affected grapes (433 and 439 µg/kg, respectively) significantly higher than the one related to healthy grapes (391 and 386 µg/kg, respectively). In response to the attack by P. japonica the plant significantly increases the content of some volatile compounds such as hexanal, (E)-2-hexenal, 1-hexanol, (E)-2-hexen-1-ol and phenyl ethyl alcohol.

Influence of Cold Pre-Fermentation Maceration on the Volatilomic Pattern and Aroma of White Wines.

Aroma compounds play a key role in wine quality due to their importance in wine aroma. The aim of the present study is to investigate the influence of cold pre-fermentative maceration (CPM) treatment on aromatic and sensory properties of white wines from four grape varieties (Tempranillo Blanco, Maturana Blanca, Viura and Garnacha Blanca) during two consecutive years (2019 and 2020). A total of 62 aroma compounds belonging to different chemical families were identified using headspace solid-phase microextraction combined with gas chromatography mass spectrometry (HS-SPME/GC-MS). CPM treatment enhanced the total relative concentration of alcohols, esters and acids compared to control wines. Regarding sensorial properties, esters made the greatest contribution to the studied white wines, mainly through the development of floral and fruity notes. On the other hand, CPM treatment did not significantly influence the total relative concentration of terpenoids, and different trends were observed according to grape variety and vintage. The obtained results showed differences in the wine's aromatic complexity according to the grape variety, the vintage and the treatment applied and suggested that CPM treatment could represent a suitable approach to manipulate the aromatic profile and enhance the aromatic quality and complexity of wine.

Behind the Scenes of Anthocyanins-From the Health Benefits to Potential Applications in Food, Pharmaceutical and Cosmetic Fields.

Anthocyanins are widespread and biologically active water-soluble phenolic pigments responsible for a wide range of vivid colours, from red (acidic conditions) to purplish blue (basic conditions), present in fruits, vegetables, and coloured grains. The pigments' stability and colours are influenced mainly by pH but also by structure, temperature, and light. The colour-stabilizing mechanisms of plants are determined by inter- and intramolecular co-pigmentation and metal complexation, driven by van der Waals, π-π stacking, hydrogen bonding, and metal-ligand interactions. This group of flavonoids is well-known to have potent anti-inflammatory and antioxidant effects, which explains the biological effects associated with them. Therefore, this review provides an overview of the role of anthocyanins as natural colorants, showing they are less harmful than conventional colorants, with several technological potential applications in different industrial fields, namely in the textile and food industries, as well as in the development of photosensitizers for dye-sensitized solar cells, as new photosensitizers in photodynamic therapy, pharmaceuticals, and in the cosmetic industry, mainly on the formulation of skin care formulations, sunscreen filters, nail colorants, skin & hair cleansing products, amongst others. In addition, we will unveil some of the latest studies about the health benefits of anthocyanins, mainly focusing on the protection against the most prevalent human diseases mediated by oxidative stress, namely cardiovascular and neurodegenerative diseases, cancer, and diabetes. The contribution of anthocyanins to visual health is also very relevant and will be briefly explored.

Bioactive Compounds and Antioxidant Activity from Spent Coffee Grounds as a Powerful Approach for Its Valorization.

Coffee is one of the world's most popular beverages, and its consumption generates copious amounts of waste. The most relevant by-product of the coffee industry is the spent coffee grounds, with 6 million tons being produced worldwide per year. Although generally treated as waste, spent coffee grounds are a rich source of several bioactive compounds with applications in diverse industrial fields. The present work aimed at the analysis of spent coffee grounds from different geographical origins (Guatemala, Colombia, Brazil, Timor, and Ethiopia) for the identification of bioactive compounds with industrial interest. For this purpose, the identification and quantification of the bioactive compounds responsible for the antioxidant activity attributed to the spent coffee grounds were attempted using miniaturized solid-phase extraction (µ-SPEed), combined with ultrahigh-performance liquid chromatography with photodiode array detection (UHPLC-PDA). After validation of the µ-SPEed/UHPLC-PDA method, this allowed us to conclude that caffeine and 5-caffeoylquinic acid (5-CQA) are the most abundant bioactive compounds in all samples studied. The total phenolic content (TPC) and antioxidant activity are highest in Brazilian samples. The results obtained show that spent coffee grounds are a rich source of bioactive compounds, supporting its bioprospection based on the circular economy concept closing the loop of the coffee value chain, toward the valorization of coffee by-products.

Spices Volatilomic Fingerprinting-A Comprehensive Approach to Explore Its Authentication and Bioactive Properties.

Volatile organic metabolites (VOMs) present in different spices can provide distinct analytical biosignatures related to organoleptic properties and health benefits. This study aimed to establish the volatilomic fingerprint of six of the most consumed spices all over the world (saffron (Crocus sativus L.), cinnamon (Cinnamomum verum), cumin (Cuminum cyminum L.), black pepper, (Piper nigrum L.), sweet paprika (Capsicum annuum L.), and curry (a mix of different herbs and spices)). Based on headspace solid phase microextraction (HS-SPME) followed by gas chromatography-mass spectrometry (GC-MS) analysis, this is a powerful strategy to explore and establish the spice's volatile pattern and unravel the potential health benefits related to the most important VOMs identified in each spice. This comprehensive knowledge will help in the definition of their authenticity, while simultaneously protecting against potential frauds and adulterations. A total of 162 VOMs were identified. Semi-quantitative assessments revealed that terpenoids and sesquiterpenoids amounted to the major volatile class in the investigated spices, except for cinnamon, where carbonyl compounds are the major group. Most of the studied spices comprised key characteristics of aroma and health bioactive compounds, e.g., dihydrojuneol in saffron, cinnamaldehyde in cinnamon, cuminaldehyde in cumin and curry, and caryophyllene in black pepper. The principal component analysis (PCA) and partial least-squares discriminant analysis (PLS-DA) successfully discriminated the investigated spices, being α-cubebene, 3-methyl butanal, ß-patchoulene and ß-selinene, the most important VOMs (highest VIP's) that contributed to its discrimination. Moreover, some VOMs have a high influence on the spice's bioactive potential, helping to prevent certain diseases including cancer, inflammatory-related diseases, diabetes, and cardiovascular diseases.

Chromatographic-Based Platforms as New Avenues for Scientific Progress and Sustainability.

Chromatography was born approximately one century ago and has undergone outstanding technological improvements in innovation, research, and development since then that has made it fundamental to advances in knowledge at different levels, with a relevant impact on the well-being and health of individuals. Chromatography boosted a comprehensive and deeper understanding of the complexity and diversity of human-environment interactions and systems, how these interactions affect our life, and the several societal challenges we are currently facing, namely those related to the sustainability of our planet and the future generations. From the life sciences, which allowed us to identify endogenous metabolites relevant to disease mechanisms, to the OMICS field, nanotechnology, clinical and forensic analysis, drug discovery, environment, and "foodprint", among others, the wide range of applications of today's chromatographic techniques is impressive. This is fueled by a great variability of powerful chromatographic instruments currently available, with very high sensitivity, resolution, and identification capacity, that provide a strong basis for an analytical platform able to support the challenging demands of the postgenomic and post COVID-19 eras. Within this context, this review aims to address the great utility of chromatography in helping to cope with several societal-based challenges, such as the characterization of disease and/or physiological status, and the response to current agri-food industry challenges of food safety and sustainability, or the monitoring of environmental contamination. These are increasingly important challenges considering the climate changes, the tons of food waste produced every day, and the exponential growth of the human population. In this context, the principles governing the separation mechanisms in chromatography as well the different types and chromatographic techniques will be described. In addition, the major achievements and the most important technological advances will be also highlighted. Finally, a set of studies was selected in order to evince the importance of different chromatographic analyses to understand processes or create fundamental information in the response to current societal challenges.

Recent Developments in the Applications of Fingerprinting Technology in the Food Field.

In recent years, the concerns and demands by consumers for the high quality and safety for natural and processed plant-based and animal foods has increased significantly [...].

Valorization of Spent Coffee Grounds as a Natural Source of Bioactive Compounds for Several Industrial Applications-A Volatilomic Approach.

Coffee is one of the most popular beverages worldwide, whose production and consumption result in large amounts of waste, namely spent coffee grounds, constituting an important source of compounds for several industrial applications. This work focused on the establishment of the volatile fingerprint of five spent coffee grounds from different geographical origins using headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry (HS-SPME/GC-MS), as a strategy to identify volatile organic metabolites (VOMs) with potential application in the food industry as antioxidant, anti-inflammatory, and antiproliferative agents. One hundred eleven VOMs belonging to different chemical families were identified, of which 60 were found in all spent coffee grounds analyzed. Furanic compounds (34%), nitrogen compounds (30%), and esters (19%) contributed significant to the total volatile fingerprint. The data obtained suggest that spent coffee grounds have great potential to be used as raw material for different approaches in the food industry towards the development of new food ingredients or products for human consumption, in addition to pharmaceutical and cosmetic applications, namely as antioxidant (e.g., limonene, carvacrol), antimicrobial (e.g., pyrrole-2-carboxaldehyde, ß-myrcene) and anti-inflammatory (e.g., furfural, 2-furanmethanol) agents, promoting their integral valorization within the circular bioeconomy concept.

High throughput analytical approach based on µQuEChERS combined with UHPLC-PDA for analysis of bioactive secondary metabolites in edible flowers.

Mallow blue (Malva sylvestris L.), hibiscus (Hibiscus rosa-sinensis L.) and nasturtium (Tropaeolum majus L.), are common edible flowers rich in bioactive secondary metabolites (BASMs) whose use in sophisticated gastronomy present currently as increasing trend. In this study the BASMs profile of these edible flowers was established using an emerging green extraction technique, µQuEChERS followed by ultra-high performance liquid chromatography coupled to a photodiode array detection system (UHPLC-PDA). After validation the µQuEChERS/UHPLC-PDA methodology allow to identify that apigenin and epigallocatechin gallate are the most abundant BASMs in mallow blue flowers, while catechin and dicaffeoylquinic acid are predominant in hibiscus flowers, and myricitrin and dicaffeoylquinic acid in nasturtium flowers. Total polyphenol content is the highest in the extract of hibiscus. Nasturtium shows the greatest radical scavenging activity. The results revealed that these flowers constitute a potential source of BASMs with different bioactive properties suggesting its use in design of new functional foods.

Green Extraction Techniques as Advanced Sample Preparation Approaches in Biological, Food, and Environmental Matrices: A Review.

Green extraction techniques (GreETs) emerged in the last decade as greener and sustainable alternatives to classical sample preparation procedures aiming to improve the selectivity and sensitivity of analytical methods, simultaneously reducing the deleterious side effects of classical extraction techniques (CETs) for both the operator and the environment. The implementation of improved processes that overcome the main constraints of classical methods in terms of efficiency and ability to minimize or eliminate the use and generation of harmful substances will promote more efficient use of energy and resources in close association with the principles supporting the concept of green chemistry. The current review aims to update the state of the art of some cutting-edge GreETs developed and implemented in recent years focusing on the improvement of the main analytical features, practical aspects, and relevant applications in the biological, food, and environmental fields. Approaches to improve and accelerate the extraction efficiency and to lower solvent consumption, including sorbent-based techniques, such as solid-phase microextraction (SPME) and fabric-phase sorbent extraction (FPSE), and solvent-based techniques (µQuEChERS; micro quick, easy, cheap, effective, rugged, and safe), ultrasound-assisted extraction (UAE), and microwave-assisted extraction (MAE), in addition to supercritical fluid extraction (SFE) and pressurized solvent extraction (PSE), are highlighted.

Comprehensive Evaluation of the Volatomic Fingerprint of Saffron from Campania towards Its Authenticity and Quality.

The volatile profiles of eight saffron samples (seven cultivated and one spontaneous) grown in different geographical districts within the Campania region (southern Italy) were compared. Using headspace solid-phase microextraction coupled to gas chromatography-mass spectrometry (HS-SPME/GC-MS), overall, 80 volatiles were identified in the eight landraces. Among them, safranal and its isomers and other related compounds such as isophorones, which are not only key odorants but also pharmacologically active metabolites, have been detected in all the investigated samples. Principal Component Analysis performed on the volatiles' compounds revealed that the spontaneous sample turned out to be an outlier. In particular, the volatile organic compounds (VOCs) profile of the spontaneous saffron presented four lilac aldehydes and four lilac alcohol isomers, which, to the authors' knowledge, have never been identified in the volatile signature of this spice. The multivariate statistical analysis allowed the discrimination of the seven cultivate saffron ecotypes in four well-separated clusters according to variety. Moreover, 20 VOCs, able to differentiate the clusters in terms of single volatile metabolite, were discovered. Altogether, these results could contribute to identifying possible volatile signature metabolites (biomarkers) or patterns that discriminate saffron samples grown in Campania region on a molecular basis, encouraging future biodiversity programs to preserve saffron landraces revealing valuable genetic resources.

Improved approach based on MALDI-TOF MS for establishment of the fish mucus protein pattern for geographic discrimination of Sparus aurata.

Food fraud is still a recurrent practice throughout food supply chains. In the case of seafood, misidentification of species and products repackaging constitute the most common frauds. Therefore, the development of appropriate analytical approaches to be used against food fraud is necessary. The present study goal is to explore for the first time, the possibility to differentiate between Sparus aurata from two different mariculture farms located in Madeira Island (Caniçal and Ribeira Brava), using the mass fingerprint of fish mucus obtained from MALDI-TOF MS and analyzed using Mass-UP software for multivariate statistical analysis and biomarker identification. It was possible to establish, from the mucus protein fraction, a set of potential biomarkers for each location in a total of 35 peaks, being 17 peaks specific to Caniçal located farm and 18 to Ribeira Brava. The proposed analytical approach revealed a useful strategy providing accurate and fast results for fish geographical origin discrimination.

Evaluation of Fatty Acids Profile as a Useful Tool towards Valorization of By-Products of Agri-Food Industry.

Worldwide, the food industry generates a large number of by-products from a wide variety of sources. These by-products represent an interesting and economical source of added value components with potential functionalities and/or bioactivities, which might be explored for industrial purposes, encouraging and promoting the circular economy concept. In this context, the current work aimed to evaluate the fatty acids (FAs) profile using gas chromatography-flame ionization detector (GC-FID) and Fourier Transform Infrared (FTIR), as well as the determination of related health lipid indices (e.g., atherogenic (AI) and thrombogenic (TI)) as a powerful strategy to investigate the potential applications of different agri-food by-products for human nutrition and animal feeding. This work results showed that polyunsaturated fatty acids (PUFAs) are the predominant group in grape pomace (72.7%), grape bunches (54.3%), and brewer's spent grain (BSG, 59.0%), whereas carrot peels are dominated by monounsaturated fatty acids (MUFAs, 47.3%), and grape stems (46.2%), lees (from 50.8 to 74.1%), and potato peels (77.2%) by saturated fatty acids (SFAs). These findings represent a scientific basis for exploring the nutritional properties of agri-food by-products. Special attention should be given to grape pomace, grape bunches, and BSG since they have a high content of PUFAs (from 54.3 to 72.7%) and lower AI (from 0.11 to 0.38) and TI (from 0.30 to 0.56) indexes, suggesting their potential to provide a variety of health benefits against cardiovascular diseases including well-established hypotriglyceridemia and anti-inflammatory effects, products to which they are added.

An integrative approach based on GC-qMS and NMR metabolomics data as a comprehensive strategy to search potential breast cancer biomarkers.

INTRODUCTION: Globally, breast cancer (BC) is leading at the top of women's diseases and, as a multifactorial disease, there is the need for the development of new approaches to aid clinicians on monitoring BC treatments. In this sense, metabolomic studies have become an essential tool allowing the establishment of interdependency among metabolites in biological samples. OBJECTIVE: The combination of nuclear magnetic resonance (NMR) and gas chromatography-quadrupole mass spectrometry (GC-qMS) based metabolomic analyses of urine and breast tissue samples from BC patients and cancer-free individuals was used. METHODS: Multivariate statistical tools were used in order to obtain a panel of metabolites that could discriminate malignant from healthy status assisting in the diagnostic field. Urine samples (n = 30), cancer tissues (n = 30) were collected from BC patients, cancer-free tissues were resected outside the tumor margin from the same donors (n = 30) while cancer-free urine samples (n = 40) where obtained from healthy subjects and analysed by NMR and GC-qMS methodologies. RESULTS: The orthogonal partial least square discriminant analysis model showed a clear separation between BC patients and cancer-free subjects for both classes of samples. Specifically, for urine samples, the goodness of fit (R2Y) and predictive ability (Q2) was 0.946 and 0.910, respectively, whereas for tissue was 0.888 and 0.813, revealing a good predictable accuracy. The discrimination efficiency and accuracy of tissue and urine metabolites was ascertained by receiver operating characteristic curve analysis that allowed the identification of metabolites with high sensitivity and specificity. The metabolomic pathway analysis identified several dysregulated pathways in BC, including those related with lactate, valine, aspartate and glutamine metabolism. Additionally, correlations between urine and tissue metabolites were investigated and five metabolites (e.g. acetone, 3-hexanone, 4-heptanone, 2-methyl-5-(methylthio)-furan and acetate) were found to be significant using a dual platform approach. CONCLUSION: Overall, this study suggests that an improved metabolic profile combining NMR and GC-qMS may be useful to achieve more insights regarding the mechanisms underlying cancer.