Study of the minor fraction of virgin olive oil by a multi-class GC-MS approach: Comprehensive quantitative characterization and varietal discrimination potential.

For the first time, a multi-class GC-MS method was applied to perform the quantitative-profiling of the minor fraction of VOOs (considering >40 compounds) in a single run. This comprehensive methodology has demonstrated a remarkable profiling ability on five groups of compounds (phenolic and triterpenic compounds, tocopherols, sterols and free fatty acids) with wide range of polarities/volatilities and chemical entities. After the complete analytical validation of the method, 32 VOO samples from eight different cultivars (some of them very scarcely studied before) were analyzed and the quantitative results were subjected to both non-supervised and supervised multivariate statistics for testing the capability of the determined VOO minor compounds to discriminate the varietal origin of the samples. Typical compositional profiles were defined for each cultivar and promising potential varietal markers were pointed out. The models built to discriminate Cayon and Maurino samples from the rest exhibited the best quality parameters. The relative levels of tocopherols together with characteristic concentration of luteolin, ß-sitosterol and tyrosol were, for instance, the most specific features of Cayon VOOs.

Characterization of New Olive Fruit Derived Products Obtained by Means of a Novel Processing Method Involving Stone Removal and Dehydration with Zero Waste Generation.

As a result of an innovative olive fruit processing method involving stone removal and dehydration, a new kind of olive oil and olive flour are generated. The main objective of this work was to accomplish the comprehensive characterization of the minor compounds of both products and to evaluate the effect of the dehydration temperature on their composition. To this end, olive oil and flour samples obtained through the novel processing method were analyzed and compared with "conventional" virgin olive oils (VOO). The applied LC-MS methodology allowed the determination of 57 metabolites belonging to different chemical classes (phenolic compounds, pentacyclic trirterpenes, and tocopherols). Both the new oils and flours presented considerable amounts of olive fruit metabolites that are usually absent from VOO. Quantitative differences were found among VOOs and the new oils, probably due to the inhibition of some enzymes caused by the temperature increase or the absence of water during the processing.

New insight into phenolic composition of chayote (Sechium edule (Jacq.) Sw.).

Chayote, the fruit of Sechium edule (Jacq.) Swartz, is known for culinary applications and in traditional medicine. A great number of its health benefits have been related to its phenolic composition. However, literature concerning the characterization of phenolic compounds in chayote is scarce. Therefore, this work highlights new information about the composition of chayote. For that, different hydro-alcoholic mixtures (50-100% ethanol content) were used to recover phenolic compounds by ultrasound assisted extraction (UAE) and the platform HPLC-ESI-TOF-MS was employed to characterize these components in the fruit. Briefly, 26 compounds, classified as phenolic acids, flavonoids and other polar compounds were identified in the fruit. About the effect of ethanol ratio on phenolic extraction, 70% ethanol exhibited the best recovery of target compounds (188 ±â€¯2 mg/100 g fruit d.w.). Chayote is rich in flavones accounting for a 60.6% from the total compounds quantified and the major compound is diosmetin 7-O-rutinoside (23.8%).

Leaf removal at veraison stage differentially affects qualitative attributes and bioactive composition of fresh and dehydrated grapes of two indigenous Cypriot cultivars.

BACKGROUND: The effect of leaf removal on postharvest performance of dehydrated grapes has been poorly analyzed. The aim of this study was to evaluate the effect of leaf removal at veraison stage on the metabolites of fresh and dehydrated grapes of two indigenous Cypriot cultivars ('Mavro' and 'Xynisteri'), which are destined for the production of 'Commandaria', a protected designation of origin (PDO) premium sweet wine. RESULTS: Leaf removal led to a reduction of soluble solids, titratable acidity, aroma potential and most of the phenolic groups in the musts of both cultivars. Dehydration led to a significant increase in all of these parameters in both cultivars, being more pronounced in cv. 'Mavro'. Interestingly, leaf removal indicated differential response in the dehydrated product of the cultivars examined; liquid chromatography (LC-DAD-qTOF-MS) data showed a significant decrease in phenolic compounds in 'Xynisteri' must (from 66.73 to 44.15 mg L-1 ), while 'Mavro' must registered similar values of phenolic compounds (from 94.78 to 96.72 mg L-1 ), but with a different distribution among phenolic groups. Intriguingly, flavonols and flavan-3-ols that present significant health-promoting properties showed higher concentrations in the must from dehydrated 'Mavro' grapes that were subjected to preharvest leaf removal. CONCLUSIONS: Leaf removal at veraison stage followed by sun drying differentially affected the chemical composition of the examined cultivars. This preharvest application followed by postharvest dehydration was beneficial for cv. 'Mavro', while this was not the case for cv. 'Xynisteri'. The effects of leaf removal in other developmental stages (i.e. pre/post bloom stage) need to be dissected with the ultimate goal of providing an end product with high bioactive content. © 2018 Society of Chemical Industry.

Evaluating the reliability of specific and global methods to assess the phenolic content of virgin olive oil: Do they drive to equivalent results?

Despite the huge number of different published methodologies, there is an open debate regarding which one is the most convenient analytical strategy for the determination of phenolic compounds from virgin olive oils. Diverse technical issues together with the disparity of criteria regarding results expression cause a lot of confusion. Herewith, a systematic comparison between specific (a powerful and LC-MS method) and global methodologies (the Folin-Ciocalteau (FC) colorimetric assay, the International Olive Council (IOC) method and hydrolysis plus HPLC-DAD) has been carried out. Thus, these strategies have been applied to the analysis of 50 extra virgin olive oils (covering all the possible quantitative ranges of these substances). This is the first time in which the individual LC-MS quantification of so many phenolic substances is included in this kind of comprehensive comparison. The outcomes of all the strategies have been thoroughly confronted and their equivalence (or divergence) has been carefully evaluated, establishing possible correspondence factors. The LC-MS individual determination with the pure standard of every analyte represented the ideal situation; when only the commercially available standards were used, a drastic change was observed in the absolute concentrations of oleuropein derivatives (in terms of hydroxytyrosol). Total phenolic content (summing individual levels) proved to be higher (1.9-3.0 times when data was expressed in mg/kg) than the values given by the three non-specific methods (with R2 from 0.84 to 0.90). In any case, the IOC method, the FC assay and the hydrolysis approach could be considered as feasible strategies when a global value is pursued. Good correlations between their results were found (R2 > 0.89), with the following equivalence factors: FC(mg caffeic acid/kg) ≈ 0.60 IOC(mg TY/kg); IOC(mg TY/kg) ≈ 1.27 Sum acid hydrolysis(mg TY+HTY/kg); FC(mg HTY/kg) ≈ 1.04 Sum acid hydrolysis(mg TY+HTY/kg).

Establishing the Phenolic Composition of Olea europaea L. Leaves from Cultivars Grown in Morocco as a Crucial Step Towards Their Subsequent Exploitation.

In Morocco, the recovery of olive agro-industrial by-products as potential sources of high-added value substances has been underestimated so far. A comprehensive quantitative characterization of olive leaves' bioactive compounds is crucial for any attempt to change this situation and to implement the valorization concept in emerging countries. Thus, the phenolic fraction of olive leaves of 11 varieties ('Arbequina', 'Hojiblanca', 'Frantoio', 'Koroneiki', 'Lechín', 'Lucque', 'Manzanilla', 'Picholine de Languedoc', 'Picholine Marocaine', 'Picual' and 'Verdal'), cultivated in the Moroccan Meknès region, was investigated. Thirty eight phenolic or related compounds (including 16 secoiridoids, nine flavonoids in their aglycone form, seven flavonoids in glycosylated form, four simple phenols, one phenolic acid and one lignan) were determined in a total of 55 samples by using ultrasonic-assisted extraction and liquid chromatography coupled to electrospray ionization-ion trap mass spectrometry (LC-ESI-IT MS). Very remarkable quantitative differences were observed among the profiles of the studied cultivars. 'Picholine Marocaine' variety exhibited the highest total phenolic content (around 44 g/kg dry weight (DW)), and logically showed the highest concentration in terms of various individual compounds. In addition, chemometrics (principal components analysis (PCA) and stepwise-linear discriminant analysis (s-LDA)) were applied to the quantitative phenolic compound data, allowing good discrimination of the selected samples according to their varietal origin.

Unravelling the Distribution of Secondary Metabolites in Olea europaea L.: Exhaustive Characterization of Eight Olive-Tree Derived Matrices by Complementary Platforms (LC-ESI/APCI-MS and GC-APCI-MS).

In order to understand the distribution of the main secondary metabolites found in Olea europaea L., eight different samples (olive leaf, stem, seed, fruit skin and pulp, as well as virgin olive oil, olive oil obtained from stoned and dehydrated fruits and olive seed oil) coming from a Picudo cv. olive tree were analyzed. All the experimental conditions were selected so as to assure the maximum coverage of the metabolome of the samples under study within a single run. The use of LC and GC with high resolution MS (through different ionization sources, ESI and APCI) and the annotation strategies within MetaboScape 3.0 software allowed the identification of around 150 compounds in the profiles, showing great complementarity between the evaluated methodologies. The identified metabolites belonged to different chemical classes: triterpenic acids and dialcohols, tocopherols, sterols, free fatty acids, and several sub-types of phenolic compounds. The suitability of each platform and polarity (negative and positive) to determine each family of metabolites was evaluated in-depth, finding, for instance, that LC-ESI-MS (+) was the most efficient choice to ionize phenolic acids, secoiridoids, flavonoids and lignans and LC-APCI-MS was very appropriate for pentacyclic triterpenic acids (MS (-)) and sterols and tocopherols (MS (+)). Afterwards, a semi-quantitative comparison of the selected matrices was carried out, establishing their typical features (e.g., fruit skin was pointed out as the matrix with the highest relative amounts of phenolic acids, triterpenic compounds and hydroxylated fatty acids, and seed oil was distinctive for its high relative levels of acetoxypinoresinol and tocopherols).

Development of a folic acid molecularly imprinted polymer and its evaluation as a sorbent for dispersive solid-phase extraction by liquid chromatography coupled to mass spectrometry.

This work shows the development of a molecularly imprinted polymer to determine folic acid (FA) in food extracts by using dispersive solid-phase extraction and liquid chromatography coupled to mass spectrometry (LC-MS). Herewith, combinations of monomers (methacrylic acid (MAA), 4-vinylpyridine (4VPy) and vinylbenzyl trimethylammonium chloride (VBTMAC)) and crosslinkers (ethylene glycol dimethacrylate (EGDMA) and divinyl benzene (DVB)) were tested in appropriate solvents. Isotherm tests revealed that the MIP with the highest affinity was obtained by combining VBTMAC and EGDMA. Having checked the appropriate template-monomer-crosslinker ratio, the FA MIP was analyzed for its kinetic and equilibrium binding properties, proving very high affinity (more than 2.5 mmol g-1) and MIP/NIP ratio (up to 37). The FA MIP was used to selectively isolate the compound of interest from lettuce and cookies matrices using a dispersive solid-phase extraction protocol (which exhibited appropriate recovery and repeatability, ≥79.50% and ≤13.41 (%RSD in terms of area values), respectively, as well as absence of matrix effect); the resulting extracts were analyzed by a rapid and reliable LC-MS method.

Evaluation of two sterically directed attachments of biomolecules on a coaxial nanofibre membrane to improve the development of optical biosensors.

In this study, we have optimised the sterically directed attachment of biomolecules on the surface of coaxial membranes prepared by co-electrospinning which have been proved to be a material with very high performance for the development of biosensors with optical oxygen transduction. Uricase has been used as model enzyme. Two sterically directed strategies: a) covalent attachment via maleimide, and b) affinity bonding via biotin-streptavidin interaction, have been tested in order to preserve the enzymatic activity of uricase and to improve the analytical figures of merits on the determination of uric acid. The best results were obtained with biotin-streptavidin affinity interaction and using a biotinylation reagent containing a polyethylene glycol chain. The developed biosensor showed high sensitivity towards uric acid with a detection limit of 0.5 µM, a quantification limit of 1.8 µM and linear range from 1.8 to 250 µM. The applicability of the membrane as biosensor with optical oxygen transduction was proved by determining uric acid in serum samples. The obtained results showed a good correlation (0.999) with those obtained by an external reference laboratory.

Deep insight into the minor fraction of virgin olive oil by using LC-MS and GC-MS multi-class methodologies.

Several analytical methods are available to evaluate virgin olive oil (VOO) minor compounds; however, multi-class methodologies are yet rarely studied. Herewith, LC-MS and GC-MS platforms were used to develop two methods capable of simultaneously determine more than 40 compounds belonging to different VOO minor chemical classes within a single run. A non-selective and highly efficient liquid-liquid extraction protocol was optimized for VOO minor components isolation. The separation and detection conditions were adjusted for determining phenolic and triterpenic compounds, free fatty acids and tocopherols by LC-MS, plus sterols and hydrocarbons by GC-MS. Chromatographic analysis times were 31 and 50 min, respectively. A comparative assessment of both methods in terms of analytical performance, easiness, cost and adequacy to the analysis of each class was carried out. The emergence of this kind of multi-class analytical methodologies greatly increases throughput and reduces cost, while avoiding the complexity and redundancy of single-chemical class determinations.

Characterization of bioactive compounds of Annona cherimola L. leaves using a combined approach based on HPLC-ESI-TOF-MS and NMR.

Annona cherimola Mill. (cherimoya) has widely been used as food crop. The leaves of this tree possess several health benefits, which are, in general, attributed mainly to its bioactive composition. However, literature concerning a comprehensive characterization based on a combined approach, which consists of nuclear magnetic resonance (NMR) and high-performance liquid chromatography coupled with time-of-flight mass spectrometry (HPLC-TOF-MS), from these leaves is scarce. Thus, the aim of this work was to study the polar profile of full extracts of cherimoya leaves by using these tools. Thus, a total of 77 compounds have been characterized, 12 of which were identified by both techniques. Briefly, 23 compounds were classified as amino acids, organic acids, carbohydrates, cholines, phenolic acid derivatives, and flavonoids by NMR, while 66 metabolites were divided into sugars, amino acids, phenolic acids and derivatives, flavonoids, phenylpropanoids, and other polar compounds by HPLC-TOF-MS. It is worth mentioning that different solvent mixtures were tested and the total phenolic content in the extracts quantified (TPC via HPLC-TOF-MS). The tendency observed was EtOH/water 80/20 (v/v) (17.0 ± 0.2 mg TPC/g leaf dry weight (d.w.)) ≥ acetone/water 70/30 (v/v) (16.1 ± 0.7 mg TPC/g leaf d.w.) > EtOH/water 70/30 (v/v) (14.0 ± 0.3 mg TPC/g leaf d.w.) > acetone/water 80/20 (v/v) (13.5 ± 0.4 mg TPC/g leaf d.w.). Importantly, flavonoids derivatives were between 63 and 76% of the TPC in those extracts. Major compounds were sucrose, glucose (α and ß), and proline, and chlorogenic acid and rutin for NMR and HPLC-TOF-MS, respectively. Graphical abstract The combined use of LC-HRMS and NMR is a potential synergic combination for a comprehensive metabolite composition of cherimoya leaves.

A multifunctional material based on co-electrospinning for developing biosensors with optical oxygen transduction.

A multifunctional material based on co-electrospinning has been developed as a basic material for the development of biosensors with optical oxygen transduction. It is based on coaxial nanofibres: inner fibres containing an oxygen sensitive dye and outer fibres containing aldehyde groups to allow the formation of Schiff bases with the amino groups of the enzyme. The resulting material preserves the oxygen sensing properties of the inner optical transducer as well as exhibits a high capacity for immobilizing molecules on its surface. Uricase has been selected as model enzyme and several parameters (temperature, pH, reaction time, buffer, and enzyme concentration) have been optimised to demonstrate the versatility of this novel multifunctional material in the development of biosensors with optical oxygen transduction for determining uric acid in serum samples. It suggests that the proposed multifunctional material can provide a promising multifunctional platform for biosensing applications.

A metabolic fingerprinting approach based on selected ion flow tube mass spectrometry (SIFT-MS) and chemometrics: A reliable tool for Mediterranean origin-labeled olive oils authentication.

Selected Ion flow tube mass spectrometry (SIFT-MS) in combination with chemometrics was used to authenticate the geographical origin of Mediterranean virgin olive oils (VOOs) produced under geographical origin labels. In particular, 130 oil samples from six different Mediterranean regions (Kalamata (Greece); Toscana (Italy); Meknès and Tyout (Morocco); and Priego de Córdoba and Baena (Spain)) were considered. The headspace volatile fingerprints were measured by SIFT-MS in full scan with H3O+, NO+ and O2+ as precursor ions and the results were subjected to chemometric treatments. Principal Component Analysis (PCA) was used for preliminary multivariate data analysis and Partial Least Squares-Discriminant Analysis (PLS-DA) was applied to build different models (considering the three reagent ions) to classify samples according to the country of origin and regions (within the same country). The multi-class PLS-DA models showed very good performance in terms of fitting accuracy (98.90-100%) and prediction accuracy (96.70-100% accuracy for cross validation and 97.30-100% accuracy for external validation (test set)). Considering the two-class PLS-DA models, the one for the Spanish samples showed 100% sensitivity, specificity and accuracy in calibration, cross validation and external validation; the model for Moroccan oils also showed very satisfactory results (with perfect scores for almost every parameter in all the cases).

Development and validation of LC-MS-based alternative methodologies to GC-MS for the simultaneous determination of triterpenic acids and dialcohols in virgin olive oil.

Pentacyclic triterpenes are minor, but very relevant compounds found in virgin olive oil (VOO). A rapid and reliable LC-MS method for determining the triterpenic acids and dialcohols (after ultrasound assisted extraction) from VOO has been developed, giving an alternative to the widely used GC (FID/MS) methodologies. The analytical parameters of the proposed method were exhaustively checked, establishing limits of detection (from 1 to 95µg/l) and quantification, precision (RSD values for inter-day repeatability were found between 4.2 and 7.3% considering area values), trueness (within the range 92.7 and 100.5%) and evaluating possible matrix effect (which was no significant). The method was applied to the analysis of six triterpenic compounds in 11 monovarietal VOOs and the results compared with the quantitative GC-MS data. Moreover, the direct injection (after a simple dilution) of the samples into the LC-MS system was also tested, in an attempt to proffer an even simpler sample treatment.

Olive oil authentication: A comparative analysis of regulatory frameworks with especial emphasis on quality and authenticity indices, and recent analytical techniques developed for their assessment. A review.

Over the last decades, olive oil quality and authenticity control has become an issue of great importance to consumers, suppliers, retailers, and regulators in both traditional and emerging olive oil producing countries, mainly due to the increasing worldwide popularity and the trade globalization of this product. Thus, in order to ensure olive oil authentication, various national and international laws and regulations have been adopted, although some of them are actually causing an enormous debate about the risk that they can represent for the harmonization of international olive oil trade standards. Within this context, this review was designed to provide a critical overview and comparative analysis of selected regulatory frameworks for olive oil authentication, with special emphasis on the quality and purity criteria considered by these regulation systems, their thresholds and the analytical methods employed for monitoring them. To complete the general overview, recent analytical advances to overcome drawbacks and limitations of the official methods to evaluate olive oil quality and to determine possible adulterations were reviewed. Furthermore, the latest trends on analytical approaches to assess the olive oil geographical and varietal origin traceability were also examined.

Use of HPLC- and GC-QTOF to determine hydrophilic and lipophilic phenols in mango fruit (Mangifera indica L.) and its by-products.

Mango industry processing generates high quantities of mango by-products such as peels and seeds (35%-60% of the fruit). Indeed, it is known that mango and its by-products contain different families of bioactive compounds that possess several health benefits. Thus, the aim of this study has been the determination of different families of phenolic derivatives (free and bound phenolic compounds and alk(en)ylresorcinols (ARs)) in mango edible part and its by-products (peel, seed and seed husk) from three different cultivars. This is the first study that evaluates the phenolic compounds and ARs in the four fractions of mango of three different cultivars. Special attention has been paid to the determination of anthocyanins and ARs, because these families of compounds had not been studied in depth in mango. In fact, petunidin rutinoside-(p-coumaric acid) gallate was found in mango pulp, peel, seed and seed husk of the three cultivars and, it had never been described in mango before. It is also important to highlight that this is the first time that the identification and quantification of ARs have been performed in mango seed and seed husk; besides, four and five out of eleven alk(en)ylresorcinols detected in peel and pulp, respectively, were identified for the first time in these mango fractions. Furthermore, antioxidant activity was measured by ABTS and FRAP assays. Seed free and bound phenolic extracts showed the highest antioxidant capacity.

Comparison of Two Stationary Phases for the Determination of Phytosterols and Tocopherols in Mango and Its By-Products by GC-QTOF-MS.

Two different gas chromatography coupled to quadrupole-time of flight mass spectrometry (GC-QTOF-MS) methodologies were carried out for the analysis of phytosterols and tocopherols in the flesh of three mango cultivars and their by-products (pulp, peel, and seed). To that end, a non-polar column ((5%-phenyl)-methylpolysiloxane (HP-5ms)) and a mid-polar column (crossbond trifluoropropylmethyl polysiloxane (RTX-200MS)) were used. The analysis time for RTX-200MS was much lower than the one obtained with HP-5ms. Furthermore, the optimized method for the RTX-200MS column had a higher sensibility and precision of peak area than the HP-5ms methodology. However, RTX-200MS produced an overlapping between ß-sitosterol and Δ5-avenasterol. Four phytosterols and two tocopherols were identified in mango samples. As far as we are concerned, this is the first time that phytosterols have been studied in mango peel and that Δ5-avenasterol has been reported in mango pulp. α- and γ-tocopherol were determined in peel, and α-tocopherol was the major tocopherol in this fraction (up to 81.2%); however, only α-tocopherol was determined in the pulp and seed. The peel was the fraction with the highest total concentration of phytosterols followed by seed and pulp, and "Sensación" was the cultivar with the highest concentration of total phytosterols in most cases. There were no significant differences between quantification of tocopherols with both columns. However, in most cases, quantification of phytosterols was higher with RTX-200MS than with HP-5ms column.

Health Effects of Psidium guajava L. Leaves: An Overview of the Last Decade.

Today, there is increasing interest in discovering new bioactive compounds derived from ethnomedicine. Preparations of guava (Psidium guajava L.) leaves have traditionally been used to manage several diseases. The pharmacological research in vitro as well as in vivo has been widely used to demonstrate the potential of the extracts from the leaves for the co-treatment of different ailments with high prevalence worldwide, upholding the traditional medicine in cases such as diabetes mellitus, cardiovascular diseases, cancer, and parasitic infections. Moreover, the biological activity has been attributed to the bioactive composition of the leaves, to some specific phytochemical subclasses, or even to individual compounds. Phenolic compounds in guava leaves have been credited with regulating blood-glucose levels. Thus, the aim of the present review was to compile results from in vitro and in vivo studies carried out with guava leaves over the last decade, relating the effects to their clinical applications in order to focus further research for finding individual bioactive compounds. Some food applications (guava tea and supplementary feed for aquaculture) and some clinical, in vitro, and in vivo outcomes are also included.

Metabolic profiling approach to determine phenolic compounds of virgin olive oil by direct injection and liquid chromatography coupled to mass spectrometry.

A LC-MS method involving direct injection of extra-virgin olive oil (EVOO) - after a simple dilution - for determining its phenolic compounds has been developed. Optimization of the most appropriate solvent for sample dilution, selection of the optimum oil/solvent ratio, and establishment of column cleaning strategy and maximum number of injections were some of the most relevant steps. Then, the analytical parameters of the method were evaluated, establishing LOD (from 3.3 to 31.6µg/L) and LOQ, precision (RSD values for inter-day repeatability were found between 3.49 and 6.12%), and trueness (within the range 89.9-102.3% for 1.0mg/L) and checking possible matrix effect (which was no significant). Three kinds of calibration were used: external standard, standard addition and calibration in a phenols-free matrix, which was subsequently applied to quantify the phenolic compounds in 16 EVOOs (from 6 cultivars). A total of 21 compounds were determined without the need of using any extraction protocol.

UHPLC/MS2-based approach for the comprehensive metabolite profiling of bean (Vicia faba L.) by-products: A promising source of bioactive constituents.

Today, by-products generated from the agro-industrial practices are considered a key source of bioactive and functional components, that can be used for their nutritional and added value properties. New aspects concerning the use of these wastes as by-products in food production additives or supplements with high nutritional and medicinal value have gained substantial interest, due to their possession of economically high-value products. In this sense, the present study describes a thorough characterization of phytochemical compounds from hydro-methanolic extract of broad beans testa by using ultra-high performance liquid chromatography (UHPLC) hyphenated with quadrupole-time-of-flight tandem mass spectrometry (QTOF-MS). The proposed analytical technique provides tentative characterization of 134 phenolic and other phytochemical compounds in the Vicia faba extract, most of which have not been described so far in broad beans. Thus, >85 phytochemicals (Amino acids, phenolic acids, flavonoids, lignans, and terpenoids derivatives) are being reported herein in broad beans pods for the first time. The characterization process was carried out using MS and MS2 data provided by the ESI-qTOF-MS, along with the use of the relevant literature based on the same botanical family. The data obtained demonstrates that the agro-industrial by-product could potentially be utilized as a promising source of bioactive ingredients to design new functional foods and nutraceuticals with a valuable future market. Furthermore, the obtained data may form a basis for future quantitative and bioavailability studies, which will be the next step in this present work.