A Bacillus-based biofungicide agent prevents ochratoxins occurrence in grapes and impacts the volatile profile throughout the Chardonnay winemaking stages.

Bacillus-based biocontrol agents have emerged as a strategy to eliminate or reduce the use of synthetic fungicides that are detrimental to health and the environment. In vineyards, a special concern arises from the control of Aspergillus carbonarius, a fungus known for its potential to produce ochratoxins. Ochratoxin A (OTA) is the most toxic form among ochratoxins and its maximum limit in wine has been established in Europe and Brazil as 2 µg/kg. Wine quality, especially the volatile profile, may be influenced by the antifungal strategies, since fungicide residues are transferred from grapes to must during winemaking. The objective of this study was to evaluate, for the first time, the impact of a biocontrol strategy containing Bacillus velezensis P1 on the volatile profile and occurrence of ochratoxins when grapes infected with A. carbonarius were used in winemaking. The evaluation of ochratoxins was carried out by liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (LC-QToF-MS), and volatile compounds were analyzed using comprehensive two-dimensional gas chromatography coupled to quadrupole mass spectrometry (GC × GC/qMS). Six ochratoxins were identified in must prepared with Chardonnay grapes inoculated with A. carbonarius (ochratoxin α, ochratoxin ß, ochratoxin α methyl-ester, ochratoxin α amide, N-formyl-ochratoxin α amide and OTA). Although winemaking causes a decrease in the levels of all forms of ochratoxins, the co-occurrence of these mycotoxins was verified in wine made with grapes containing A. carbonarius. B. velezensis P1 prevented the occurrence of ochratoxins in must, ensuring the safety of wines. Regarding the volatile profile, a predominant presence of terpenic compounds was verified in samples treated with B. velezensis when compared with those not treated with the biocontrol strategy, whereas the presence of A. carbonarius resulted in a higher concentration of volatile compounds with an odor described as fatty/waxy, possibly compromising wine quality. Therefore, B. velezensis P1 is a new biofungicide possibility to produce ochratoxin-free grapes and high-quality wines.

Comprehensive two-dimensional liquid chromatography-based quali-quantitative screening of aqueous phases from pyrolysis bio-oils.

Pyrolysis processes are an alternative to minimize the environmental problem associated to agrifood industrial wastes. The main product resulting from these processes is a high-value liquid product, called bio-oil. Recently, the use of comprehensive two-dimensional liquid chromatography (LC × LC) has been demonstrated as a useful tool to improve the characterization of the water-soluble phases of bio-oils, considering their complexity and high water content. However, the precise composition of bio-oils from different agrifood byproducts is still unknown. In the present study, the qualitative and quantitative screening of eight aqueous phases from different biomasses, not yet reported in the literature, using LC × LC is presented. The two-dimensional approach was based on the use of two reverse phase separations. An amide column in the first dimension together with a C18 column in the second dimension were employed. Thanks to the use of diode array and mass spectrometry detection, 28 compounds were identified and quantified in the aqueous phase samples with good figures of merit. Samples showed a distinct quali-quantitative composition and a great predominance of compounds belonging to aldehydes, ketones and phenols, most of them with high polarity.

Polyphenolic compounds with biological activity in guabiroba fruits (Campomanesia xanthocarpa Berg.) by comprehensive two-dimensional liquid chromatography.

This study aimed to evaluate the polyphenolic composition along with the biological activity of guabiroba (Campomanesia xanthocarpa Berg.) fruits using comprehensive two-dimensional liquid chromatography (LC × LC). A simplex centroid design comprising three solvents (methanol, 2% acetic acid, and acetonitrile) was used to optimize the extraction mixture for polyphenols from ripe and unripe guabiroba fruits. A quantitative LC × LC platform was proposed to characterize the guabiroba extracts using a RP-Amide column and a C18 column in the first and second dimensions, respectively. Antidiabetic properties, using in vitro enzyme assay models and in vivo antioxidant activity with the eukaryote model Saccharomyces cerevisiae, was measured. Total phenolics compounds were more efficiently extracted with 2% acetic acid solution and acetonitrile (50:50, v/v). A total of 37 different compounds were identified and quantified using the proposed LC × LC method (linearity ranging from 0.9990 to 0.9994, intra- and interday precision from 0.40 to 10.57% and, accuracy from 81.89 to 108.98%). Significant differences were observed between ripe and unripe guabiroba fruits, especially for the compounds geraldone and methyl galangin isomer. Guabiroba fruits showed significant antidiabetic and antioxidant properties and may be potentially adopted as part of dietary strategies in the management of early stages of type 2 diabetes and associated complications.

Green Extraction Approaches for Carotenoids and Esters: Characterization of Native Composition from Orange Peel.

Orange peel is a by-product produced in large amounts that acts as a source of natural pigments such as carotenoids. Xanthophylls, the main carotenoid class found in citrus fruit, can be present in its free form or esterified with fatty acids, forming esters. This esterification modifies the compound's chemical properties, affecting their bioavailability in the human body, and making it important to characterize the native carotenoid composition of food matrices. We aimed to evaluate the non-saponified carotenoid extracts of orange peel (cv. Pera) obtained using alternative green approaches: extraction with ionic liquid (IL), analyzed by high performance liquid chromatography coupled to a diode array detector with atmospheric pressure chemical ionization and mass spectrometry HPLC-DAD-APCI-MS, and supercritical fluid extraction (SFE), followed by supercritical fluid chromatography with atmospheric pressure chemical ionization and triple quadrupole mass spectrometry detection (SFC-APCI/QqQ/MS) in an online system. Both alternative green methods were successfully applied, allowing the total identification of five free carotenoids, one apocarotenoid, seven monoesters, and 11 diesters in the extract obtained with IL and analyzed by HPLC-DAD-APCI-MS, and nine free carotenoids, six carotenoids esters, 19 apocarotenoids, and eight apo-esters with the SFE-SFC-APCI/QqQ/MS approach, including several free apocarotenoids and apocarotenoid esters identified for the first time in oranges, and particularly in the Pera variety, which could be used as a fruit authenticity parameter.

Role of the flavonoid-rich fraction in the antioxidant and cytotoxic activities of Bauhinia forficata Link. (Fabaceae) leaves extract.

Bauhinia forficata Link. is utilised as an antidiabetic in Brazilian folk-medicine; furthermore, its antioxidant properties suggest a potential usefulness in the prevention of diabetes complications associated with oxidative stress. The contribution of a flavonoid-rich fraction (FRF), HPLC-PDA-ESI-MS characterised, to the antioxidant and cytotoxic properties of B. forficata hydro-alcoholic leaves extract was evaluated for the first time. Both extract and FRF showed radical-scavenging activity and reducing power with a strong relationship with the flavonoid content found; hence, flavonoids are mainly responsible for the primary antioxidant activity of B. forficata extract. The extract significantly decreased FO-1 cell viability at the higher concentrations. FRF did not exert any effect; thus, flavonoids do not appear to be responsible for the cytotoxicity of the extract. The extract resulted virtually non-toxic against both Artemia salina and normal human lymphocytes, demonstrating potential selectivity in inhibiting cancer cell growth. Finally, no antimicrobial activity was observed against the bacteria and yeasts tested.

Native carotenoids composition of some tropical fruits.

Many tropical fruits can be considered a reservoir of bioactive substances with a special interest due to their possible health-promoting properties. The interest in carotenoids from a nutritional standpoint has recently greatly increased, because of their important health benefits. Here we report the native carotenoids composition in six tropical fruits from Panama, which is considered a region of great biodiversity. The native carotenoid composition was directly investigated by an HPLC-DAD-APCI-MS methodology, for the first time. In Corozo 32 different carotenoids were detected, including a high content of ß-carotene and lycopene. Sastra showed the highest content of zeaxanthin among the fruit investigated. In Sapote 22 different carotenoids were detected, including ß-carotene and 10 different zeaxanthin-di-esters. Frutita showed a very high content of the apo-carotenoid ß-citraurin, and of a number of its esters. In Maracuyà chino 14 carotenoids were detected, including a high amounts of mono-esterified lauric acid with ß-cryptoxanthin and with cryptocapsin. Mamey rojo was characterised by ketocarotenoids with κ rings, both hydroxylated and not hydroxylated.

Enantiomeric distribution of key volatile components in Citrus essential oils

Citrus as many other plants present characteristic distribution of some enantiomers, thus it is often possible to use this parameter for identification, characterization, genuineness, and pharmacological activity assessment. In particular, it is possible to reveal adulteration of different nature, such as addition of synthetic compounds, or natural components of different botanical origin, with drastic changes in the biological and olfactory properties. This study is focused on the evaluation of the enantiomeric excesses of numerous samples of different Citrus species: C. deliciosa Ten., C. limon (L.) Burm., C. bergamia, C. aurantifolia (Christm.) Swing., C. latifolia Tan., C. sinensis (L.) Osbeck, and C. aurantium L. The enantiomeric distribution is determined by direct esGC and, depending on the complexity of the essential oil, by MDGC with a chiral column in the second dimension. The research is focused on the determination of fourteen chiral components which present specific distribution in the essential oils investigated. Particular attention is given to the trend of the enantiomeric distribution during the productive season, so to identify useful parameters for quality assessment also in consideration of the wide range of variability often reported in literature. The components investigated were the following: α-thujene, α-pinene, camphene, β-pinene, sabinene, α-phellandrene, β-phellandrene, limonene, linalool, camphor, citronellal, linalyl acetate, terpinen-4-ol, α-terpineol. The use of MDGC allowed the separation of the enantiomers of camphor and citronellal, otherwise not separated by conventional esGC; however for the separation of the enantiomers of α-pinene it was preferable to use conventional esGC. The MDGC system allowed to determine the enantiomeric distribution of camphene, α- and β-phellandrene in lime essential oil for the first time. The results are discussed in function of seasonal variation and, when possible, in function of the extraction technology, with particular regards to lime oils.