Untargeted metabolomics of purple and orange-fleshed sweet potatoes reveals a large structural diversity of anthocyanins and flavonoids

Anthocyanins are economically valuable phytochemicals of significant relevance to human health. Industrially extracted from multiple fruit and vegetable sources, anthocyanin yield and profiles can vary between sources and growing conditions. In this study, we focused on three purple-fleshed and one orange-fleshed cultivars of sweet potato—a warm-weather, nutritious crop of substantial interest to growers in northern, cooler latitudes—to determine the yield and diversity of anthocyanins and flavonoids. Acidified ethanol extraction of lyophilized roots yielded ~ 800 mg average anthocyanins/100 g dry weight from all three cultivars. UHPLC-DAD-Orbitrap analysis of sweet potato extracts identified 18 high-confidence, mostly acylated peonidin and cyanidin derivatives contributing to > 90% of the total anthocyanin signal. Further assessment of the untargeted Liquid Chromatography–Tandem Mass Spectrometry data using deep learning and molecular networking identified over 350 flavonoid peaks with variable distributions in different sweet potato cultivars. These results provide a novel insight into anthocyanin content of purple-fleshed sweet potatoes grown in the northern latitudes, and reveal the large structural diversity of anthocyanins and flavonoids in this popular crop.

Chemical Composition, In Vitro Bioaccessibility and Antioxidant Activity of Polyphenolic Compounds from Nutraceutical Fennel Waste Extract

Fennel (Foeniculum vulgare Mill.) waste contains a broad range of bioactive molecules, including polyphenols, which have poor bioaccessibility during gastrointestinal digestion. This work aimed to investigate the bioaccessibility of total phenolic compounds and the antioxidant capacity during simulated gastrointestinal digestion using two nutraceutical formulations based on non-acid-resistant (NAR) and acid-resistant (AR) capsules containing aqueous-based extracts from fennel waste. Moreover, to obtain a comprehensive investigation of the polyphenolic constituents of the fennel waste extract, a high-resolution mass spectrometry (Q-Orbitrap) analysis was performed. Notably, chlorogenic acids, such as 4-caffeoylquinic acid and 3,4-dicaffeoylquinic acid, were the most detected compounds found in assayed samples (1.949 and 0.490 mg/g, respectively). After in vitro gastrointestinal digestion, the extract contained in AR capsules displayed higher bioaccessibility in both the duodenal and colonic stages (1.96 and 5.19 mg GAE/g, respectively) than NAR capsules (1.72 and 3.50 mg GAE/g, respectively), suggesting that the acidic gastric conditions negatively affected the polyphenol compounds released from the NAR capsules. Therefore, the aqueous extract of fennel waste could be proposed as an innovative and easily available source of dietary polyphenols. Furthermore, the use of an AR capsule could improve the polyphenol bioaccessibility and can be proposed as a nutraceutical formulation.

Antibiotics and Non-Targeted Metabolite Residues Detection as a Comprehensive Approach toward Food Safety in Raw Milk

Antibiotic residues in milk are a serious health and technological problem in dairy processes. This study aims to verify the absence of administered antimicrobials after therapeutic treatments, taking into consideration the withdrawal period, and to evaluate the reliability of screening tests under field conditions after confirmatory HPLC-HRMS (High Performance Liquid Chromatography-High-Resolution Mass Spectrometry) Orbitrap analysis. Moreover, the presence of expected or non-targeted metabolites was investigated using the new Compound Discoverer approach. The presence of antimicrobial drugs was shown in 29% of the samples, and also sometimes their metabolites (for enrofloxacin and lincomycin), despite the fact that samples were collected at the seventh milking. Moreover, in 9% of the samples, undeclared treatments were revealed due to the presence of both parent drugs and metabolites. Lastly, the putative identification of two new enrofloxacin metabolites, ENRO-N-methylacetamide and ENRO-ornithine, was proposed. In the light of this evidence, it must be borne in mind that metabolites, some of which are pharmacologically active, may also pose a risk to consumers and for the entire processing of milk in the cheese industries.

Exploring the Efficiency of UHPLC-Orbitrap MS for the Determination of 20 Pharmaceuticals and Acesulfame K in Hospital and Urban Wastewaters with the Aid of FPSE

Aside from the classical residues of persistent organic pollutants (POPs), the occurrence of emerging contaminants (ECs) in the environment has become a subject of increasing concern due to their harmful impact on the aquatic environment. Wastewater treatment plant (WWTP) effluents are major sources of environmental pollution. Therefore, data concerning their existence is required. In this study, twenty compounds representative of different drug groups considered ECs and belonging to antibiotics, antipsychotics, anti-inflammatory drugs plus acesulfame K were selected to be accurately detected and quantified with UHPLC–LTQ-Orbitrap MS in hospital and urban WWTP effluents. Chromatographic parameters (column efficiency, mobile phase, etc.), as well as mass spectrometry conditions concerning ionization mode and Orbitrap analysis (ESI options, mass resolving power, AGC target, tube lens, injection time), were evaluated. Moreover, a novel fabric phase sorptive extraction (FPSE) method based on fiber glass coated with PEG300 was employed as sample preparation process. Experimental parameters affecting extraction and desorption steps such as sample pH, extraction time, ionic strength, elution time and solvent have been optimized. The optimized methodology was validated providing excellent linearity (R2 > 0.99), and low detection and quantification limits up to 3.1 and 9.3 ng/L, for carbamazepine, respectively. Relative recoveries ranged from 81.1% to 114.0%, while a medium matrix effect for most of the target compounds occurred. Applying the above analytical method in effluents of WWTPs from NW Greece, nine compounds were quantified with concentrations that varied from 55.4 to 728.4 ng/L.

Identification of Lysine Acetylation Sites on MERS-CoV Replicase pp1ab

MERS is a life-threatening disease and MERS-CoV has the potential to cause the next pandemic. Protein acetylation is known to play a crucial role in host response to viral infection. Acetylation of viral proteins encoded by other RNA viruses have been reported to affect viral replication. It is therefore of interest to see whether MERS-CoV proteins are also acetylated. Viral proteins obtained from infected cells were trypsin-digested into peptides. Acetylated peptides were enriched by immunoprecipitation and subject to nano-LC-Orbitrap analysis. Bioinformatic analysis was performed to assess the conservation level of identified acetylation sites and to predict the upstream regulatory factors. A total of 12 acetylation sites were identified from 7 peptides, which all belong to the replicase polyprotein pp1ab. All identified acetylation sites were found to be highly conserved across MERS-CoV sequences in NCBI database. Upstream factors, including deacetylases of the SIRT1 and HDAC families as well as acetyltransferases of the TIP60 family, were predicted to be responsible for regulating the acetylation events identified. Western blotting confirms that acetylation events indeed occur on pp1ab protein by expressing NSP4 in HEK293 cells. Acetylation events on MERS-CoV viral protein pp1ab were identified for the first time, which indicate that MERS-CoV might use the host acetylation machinery to regulate its enzyme activity and to achieve optimal replication. Upstream factors were predicted, which might facilitate further analysis of the regulatory mechanism of MERS-CoV replication.