Development and Validation of a Reversed-Phase HPLC Method with UV Detection for the Determination of L-Dopa in Vicia faba L. Broad Beans

L-Dopa (LD), a substance used medically in the treatment of Parkinson's disease, is found in several natural products, such as Vicia faba L., also known as broad beans. Due to its low chemical stability, LD analysis in plant matrices requires an appropriate optimization of the chosen analytical method to obtain reliable results. This work proposes an HPLC-UV method, validated according to EURACHEM guidelines as regards linearity, limits of detection and quantification, precision, accuracy, and matrix effect. The LD extraction was studied by evaluating its aqueous stability over 3 months. The best chromatographic conditions were found by systematically testing several C-18 stationary phases and acidic mobile phases. In addition, the assessment of the best storage treatment of Vicia faba L. broad beans able to preserve a high LD content was performed. The best LD determination conditions include sun-drying storage, extraction in HCl 0.1 M, chromatographic separation with a Discovery C-18 column, 250 x 4.6 mm, 5 mu m particle size, and 99% formic acid 0.2% v/v and 1% methanol as the mobile phase. The optimized method proposed here overcomes the problems linked to LD stability and separation, thus contributing to the improvement of its analytical determination

Exploiting the anti-inflammatory potential of white capsicum extract by the nanoformulation in phospholipid vesicles

The peppers of the Capsicum species are exploited in many fields, as flavoring agents in food industry, or as decorative and therapeutic plants. Peppers show a diversified phytochemical content responsible for different biological activities. Synergic activity exerted by high levels of antioxidant compounds is responsible for their important anti-inflammatory property. A methanolic extract was obtained from a new pepper genotype and tested for anti-inflammatory activity. The extract was incorporated into phospholipid vesicles to increase the bioavailability of its bioactive components. Two types of phospholipid vesicles were produced, conventional liposomes and Penetration Enhancer containing Vesicles (PEVs). They were tested in human monoblastic leukemia U937 cell line, showing no cytotoxic effect. The intracellular reactive oxygen species (ROS) and nitric oxide (NO) levels were measured to value the in vitro efficacy of the vesicles in regulating inflammatory responses. Liposomal incorporation significantly reduced ROS levels in extract-treated LPS-activated cells. Furthermore, LC-MS/MS analyses demonstrated that liposomes facilitated the transport of the extract components across the cell membrane and their accumulation into the cytoplasm

An Interplay between a Face-Centred Composite Experimental Design and Solid-Phase Microextraction for Wine Aroma GC/MS Analysis

For oenological products, most of the intrinsic and extrinsic drivers of perceived quality are associated with specific aromatic profiles. Aromatic diversity has been recognized as a central element in perceived quality as it is able to transmit the complex interactions between grape variety, geographical characteristics, and viticultural and winemaking practices, including the fermentative process. A comprehensive characterization of flavour compounds by headspace solid-phase microextraction (HS-SPME) and gas chromatography coupled to mass spectrometric analysis is often needed in order to ascertain the quality of wine. HS-SPME requires a proper optimization that can be achieved through an adequate experimental design. Here, a HS-SPME/GC-MS based method was developed to investigate the volatile compounds of wine samples obtained by laboratory-scale fermentations. This was performed by inoculating a commercial Saccharomyces cerevisiae strain, which is used both as single starter and as mixed starter, with an indigenous Hanseniaspora osmophila strain. The experimental conditions of HS-SPME (extraction temperature and time) were optimized by applying a face-centred composite experimental design. Up to 95% of the total variance was explained by the proposed model. The optimized method allowed us to confirm the usefulness of combining the inoculation of grapes with selected yeast strains in co-culture situations in order to improve the wine bouquet

Natural polymeric materials: A solution to plastic pollution from the agro-food sector

Conventional petroleum-derived plastics represent a serious problem for global pollution because, when discarded in the environment, are believed to remain for hundreds of years. In order to reduce dependence on fossil resources, bioplastic materials are being proposed as safer alternatives. Bioplastics are bio-based and/or biodegradable materials, typically derived from renewable sources. Food waste as feedstock represents one of the recent applications in the research field of bioplastics production. To date, several food wastes have been used as raw materials for the production of bioplastics, including mostly fruit and vegetable wastes. The conversion of fruit and vegetable wastes into biomaterials could occur through simple or more complex processes. In some cases, biopolymers extracted from raw biomass are directly manufactured; on the other hand, the extracted biopolymers could be reinforced or used as reinforcing agents and/or natural fillers in order to obtain biocomposites. The present review covers available results on the application of methods used in the last 10 years for the design of biomaterials obtained from formulations made up with both fruits and vegetables by-products. Particular attention will be addressed to the waste pre-treatment, to the bioplastic formulation and to its processing, as well as to the mechanical and physical properties of the obtained materials

An Overview of Methods for L-Dopa Extraction and Analytical Determination in Plant Matrices

L-dopa is a precursor of dopamine used as the most effective symptomatic drug treatment for Parkinson’s disease. Most of the L-dopa isolated is either synthesized chemically or from natural sources, but only some plants belonging to the Fabaceae family contain significant amounts of L-dopa. Due to its low stability, the unambiguous determination of L-dopa in plant matrices requires appropriate technologies. Several analytical methods have been developed for the determination of L-dopa in different plants. The most used for quantification of L-dopa are mainly based on capillary electrophoresis or chromatographic methods, i.e., high-performance liquid chromatography (HPLC), coupled to ultraviolet-visible or mass spectrometric detection. HPLC is most often used. This paper aims to give information on the latest developments in the chemical study of L-dopa, emphasizing the extraction, separation and characterization of this compound by chromatographic, electrochemical and spectral techniques. This study can help select the best possible strategy for determining L-dopa in plant matrices using advanced analytical methods

Characterization of quercetin derivatives in a crossing combination of Habanero white and Capsicum annuum peppers and evaluation of the encapsulation efficiency of peppers extract in liposomes by LC-MS/MS

Common peppers (Capsicum) are an important source of phytochemicals, including polyphenols, flavonoids and bioactive capsaicinoids [1]. Capsaicinoids are responsible for the hot, spicy flavour presented by chilli peppers. Due to the broad range of biological activities (antioxidant, anti-inflammatory, antimicrobic, anticancer, cardioprotective) covered by Capsicum metabolites, many peppers-based nutraceutical products are available on the market [2]. Recently, novel delivery systems for enhancing the oral bioavailability of bioactive molecules are being evaluated, such as liposomes, micelles, micro/nano-emulsions, colloidal capsules and solid nanoparticles [3]. In this study, we investigated the liposomal encapsulation of a methanolic extract of white chilli peppers, obtained from an original crossing combination between Capsicum chinense (also known as Habanero white) and Capsicum annum species. An LC-MS/MS method was developed to conduct a preliminary screening of the metabolic profile of the white peppers. In addition to capsaicinoids, a deep qualitative profile of the quercetin derivatives was provided. Nine glycoconjugates were identified, i.e. quercetin-6-C-hexoside-8-C-pentoside/apioside, quercetin-4′-rhamnoside-7-hexoside, quercetin 7-O-pentoside/apioside, quercetin-3-O-pentosyl/apiosyl-rhamnosyl-hexoside, quercetin 7-O-glucoside, quercetin-3-O-rhamnoside-7-O-hexoside, quercetin-3-O-rutinose, quercetin 7-O-rhamnoside and quercetin rhamnoside-(feruloyl-hexoside). The developed LC-MS/MS method was applied to quantify the liposomial encapsulation efficiency of the white peppers extract. High encapsulation efficiency values were obtained

Untargeted metabolomic analysis by ultra-high-resolution mass spectrometry for the profiling of new Italian wine varieties

The chemical composition of wine samples comprises numerous bioactive compounds responsible for unique flavor and health-promoting properties. Thus, it’s important to have a complete overview of the metabolic profile of new wine products in order to obtain peculiar information in terms of their phytochemical composition, quality, and traceability. To achieve this aim, in this work, a mass spectrometry-based phytochemical screening was performed on seven new wine products from Villa D’Agri in the Basilicata region (Italy), i.e., Aglianico Bianco, Plavina, Guisana, Giosana, Malvasia ad acino piccolo, Colata Murro and Santa Sofia. Ultra-high-resolution mass spectrometry data were processed into absorption mode FT-ICR mass spectra, in order to remove artifacts and achieve a higher resolution and lower levels of noise. Accurate mass-to-charge ratio (m/z) values were converted into putative elemental formulas. Therefore, 2D van Krevelen diagrams were used as a tool to obtain molecular formula maps useful to perform a rapid and more comprehensive analysis of the wine sample metabolome. The presence of important metabolite classes, i.e., fatty acid derivatives, amino acids and peptides, carbohydrates and phenolic derivatives, was assessed. Moreover, the comparison of obtained metabolomic maps revealed some differences among profiles, suggesting their employment as metabolic fingerprints. This study shed some light on the metabolic composition of seven new Italian wine varieties, improving their value in terms of related bioactive compound content. Moreover, different metabolomic fingerprints were obtained for each of them, suggesting the use of molecular maps as innovative tool to ascertain their unique metabolic profile. Graphical abstract: [Figure not available: see fulltext.

Profiling of quercetin glycosides and acyl glycosides in sun-dried peperoni di Senise peppers (Capsicum annuum L.) by a combination of LC-ESI(-)-MS/MS and polarity prediction in reversed-phase separations

Interest in targeted profiling of quercetin glycoconjugates occurring in edible foodstuffs continues to expand because of their recognized beneficial health effects. Quercetin derivatives encompass several thousands of chemically distinguishable compounds, among which there are several compounds with different glycosylations and acylations. Since reference standards and dedicated databases are not available, the mass spectrometric identification of quercetin glycoconjugates is challenging. A targeted liquid chromatography (LC) coupled with tandem mass spectrometry (MS/MS) was applied for screening quercetin glycoconjugates in edible peperoni di Senise peppers (Capsicum annuum L.), protected by the European Union with the mark PGI (i.e., Protected Geographical Indication), and cultivated in Basilicata (Southern Italy). Chromatographic separation was accomplished by reversed-phase liquid chromatography (RPLC) using water/acetonitrile as the mobile phase and detection was performed on a linear ion trap mass spectrometer fitted with an electrospray ionization (ESI) source operating in negative ion mode. A correlation between experimental RP chromatographic retention time and those predicted by partition coefficients (log P) along with MS/MS data and an in-house developed database (named QUEdb) provided deep coverage for sixteen quercetin glycoconjugates. Among them, eleven quercetin glycoconjugates were already described in the literature and five were reported for the first time. These last acyl glycosidic quercetin derivatives were tentatively identified as quercetin-(galloyl-rhamnoside)-hexoside, [C34H33O20]− at m/z 761.1; quercetin-(sinapoyl-hexoside)-rhamnoside, [C38H39O20]− at m/z 815.4; quercetin-(galloyl-caffeoyl-hexoside)-rhamnoside, [C43H39O23]− at m/z 923.0; quercetin-(feruloyl-hexoside)-rhamnoside, [C37H37O19]− at m/z 785.1; and quercetin-(succinyl-rhamnoside)-rhamnoside, [C31H33O18]− at m/z 693.1. [Figure not available: see fulltext.

Metabolic profiling of Peperoni di Senise PGI bell peppers with ultra-high resolution absorption mode Fourier transform ion cyclotron resonance mass spectrometry

Bell peppers (genus Capsicum) are an excellent source of health-related compounds, such as ascorbic acid (vitamin C), carotenoids (provitamin A), tocopherols (vitamin E), flavonoids and capsaicinoids. These compounds are known for their biological activities, including antioxidant, anti-inflammatory and anticarcinogenic properties. Over the past decade, there has been an increasing number of studies focused on metabolite profiling of bell peppers. However, none of the direct analysis methods exploited in these studies were capable of simultaneously detecting compounds belonging to more than four metabolite classes (and possible derivatives of model compounds) present in bell peppers, i.e. mainly amino acids, sugars, polyphenols and organic acids. We have attempted to profile a higher number of metabolite classes by performing a direct-injection analysis with a Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS), on Peperoni di Senise bell peppers (Capsicum annuum L.). These bell peppers are a typical food product cultivated in Basilicata (Southern Italy), protected with a protected geographical indication (PGI) quality mark and known for their unique taste. To increase the performance of the FT-ICR MS method, we processed the recorded time-domain ion signals (transients) into the absorption mode FT-ICR mass spectra. The latter was achieved by the use of the dedicated software package, AutoVectis Pro. Mass spectral representation in absorption mode produced a more comprehensive description of the metabolic profile of Peperoni di Senise PGI by improving results reliability via artifacts removal and identifying a higher number of compounds

Untargeted metabolomic analysis by ultra-high-resolution mass spectrometry for the profiling of new Italian wine varieties

The chemical composition of wine samples comprises numerous bioactive compounds responsible for unique flavor and health-promoting properties. Thus, it’s important to have a complete overview of the metabolic profile of new wine products in order to obtain peculiar information in terms of their phytochemical composition, quality, and traceability. To achieve this aim, in this work, a mass spectrometry-based phytochemical screening was performed on seven new wine products from Villa D’Agri in the Basilicata region (Italy), i.e., Aglianico Bianco, Plavina, Guisana, Giosana, Malvasia ad acino piccolo, Colata Murro and Santa Sofia. Ultra-high-resolution mass spectrometry data were processed into absorption mode FT-ICR mass spectra, in order to remove artifacts and achieve a higher resolution and lower levels of noise. Accurate mass-to-charge ratio (m/z) values were converted into putative elemental formulas. Therefore, 2D van Krevelen diagrams were used as a tool to obtain molecular formula maps useful to perform a rapid and more comprehensive analysis of the wine sample metabolome. The presence of important metabolite classes, i.e., fatty acid derivatives, amino acids and peptides, carbohydrates and phenolic derivatives, was assessed. Moreover, the comparison of obtained metabolomic maps revealed some differences among profiles, suggesting their employment as metabolic fingerprints. This study shed some light on the metabolic composition of seven new Italian wine varieties, improving their value in terms of related bioactive compound content. Moreover, different metabolomic fingerprints were obtained for each of them, suggesting the use of molecular maps as innovative tool to ascertain their unique metabolic profile. Graphical abstract: [Figure not available: see fulltext.