This study was performed to assess the changes in meat quality and metabolome profiles of meat exudate during postmortem aging. At 24 h postmortem, longissimus lumborum muscles were collected from 10 pork carcasses, cut into three sections, and randomly assigned to three aging period groups (2, 9, and 16 d). Meat quality and chemical analyses, along with the metabolomics of meat exudates using ultra-high-performance liquid chromatography coupled with a quadrupole time-of-flight mass spectrometer (UHPLC-QTOF-MS) platform, were conducted. Results indicated a declined (p < 0.05) display color stability, and increased (p < 0.05) purge loss, meat tenderness, and lipid oxidation as aging extended. The principal component analysis and hierarchical clustering analysis exhibited distinct clusters of the exudate metabolome of each aging treatment. A total of 39 significantly changed features were tentatively identified via matching them to METLIN database according to their MS/MS information. Some of those features are associated with adenosine triphosphate metabolism (creatine and hypoxanthine), antioxidation (oxidized glutathione and carnosine), and proteolysis (dipeptides and tripeptides). The findings provide valuable information that reflects the meat quality’s attributes and could be used as a source of potential biomarkers for predicting aging times and meat quality changes.

Multiple reaction monitoring (MRM) profiling is a strategy for the exploratory analysis of small molecules and lipids by direct sample injection, ie, without the use of chromatographic separation. It is based on instrument methods that comprise a list of ion transitions (MRMs), in which the precursor ion is the expected ionized m/z of the lipid at its species level, ie, the description of lipid class and number of carbon and double bonds in the fatty acid chain(s), and the product ion is a fragment expected for the lipid class or for the fatty acid neutral loss. The Lipid Maps database is expanding constantly, and therefore the MRM-profiling methods associated with this database need to be continuously updated. Here, we provide a comprehensive overview and the key references for the MRM-profiling methodology and workflow, followed by a step-by-step approach to build MRM-profiling instrument acquisition methods for class-based lipid exploratory analysis based on the Lipid Maps database. The detailed workflow includes (1) importing the list of lipids from the database; (2) for a given class, combining isomeric lipids described at full structural level into 1 entry to obtain the neutral mass at species level; (3) attributing the standard Lipid Maps abbreviated nomenclature for the lipid at its species level; (4) predicting the ionized precursor ions; and (5) adding the expected product ion. We also describe how to simulate the precursor ion for the suspect screening of modified lipids using lipid oxidation and their expected product ions as an example. After determining the MRMs, information about collision energy, dwell time, and other instrument parameters are added to finalize the acquisition method. As an example of final method output, we describe the format for Agilent MassHunter v.B.06 and provide the parameters in which optimization can be performed by lipid class using one or more lipid standards.

The objective of this study was to characterize and compare the dry-aging flavor precursors and their liberation mechanisms in beef aged with different methods. Thirteen paired loins were collected at 5 days postmortem, divided into four sections, and randomly assigned into four aging methods (wet-aging (WA), conventional dry-aging (DA), dry-aging in a water-permeable bag (DWA), and UV-light dry-aging (UDA)). All sections were aged for 28 days at 2 °C, 65% RH, and a 0.8 m/s airflow before trimming and sample collection for chemical, metabolomics, and microbiome analyses. Higher concentrations of free amino acids and reducing sugars were observed in all dry-aging samples (p < 0.05). Similarly, metabolomics revealed greater short-chain peptides in the dry-aged beef (p < 0.05). The DWA samples had an increase in polyunsaturated free fatty acids (C18:2trans, C18:3n3, C20:2, and C20:5; p < 0.05) along with higher volatile compound concentrations compared to other aging methods (aldehyde, nonanal, octanal, octanol, and carbon disulfide; p < 0.05). Microbiome profiling identified a clear separation in beta diversity between dry and wet aging methods. The Pseudomonas spp. are the most prominent bacterial species in dry-aged meat, potentially contributing to the greater accumulation of flavor precursor concentrations in addition to the dehydration process during the dry-aging. Minor microbial species involvement, such as Bacillus spp., could potentially liberate unique and potent flavor precursors.

The objective of this study was to evaluate the effects of photoperiod on meat quality, oxidative stability, and metabolites of broiler fillet (M. Pectoralis major) muscles. A total of 432 broilers was split among 4 photoperiod treatments [hours light(L):dark(D)]: 20L:4D, 18L:6D, 16L:8D, and 12L:12D. At 42 days, a total of 48 broilers (12 broilers/treatment) was randomly selected and harvested. At 1 day postmortem, fillet muscles were dissected and displayed for 7 days. No considerable impacts of photoperiods on general carcass and meat quality attributes, such as carcass weight, yield, pH, water-holding capacity, and shear force, were found (p > 0.05). However, color and oxidative stability were influenced by the photoperiod, where muscles from 20L:4D appeared lighter and more discolored, coupled with higher lipid oxidation (p < 0.05) and protein denaturation (p = 0.058) compared to 12L:12D. The UPLC–MS metabolomics identified that 20 metabolites were different between the 20L:4D and 12L:12D groups, and 15 were tentatively identified. In general, lower aromatic amino acids/dipeptides, and higher oxidized glutathione and guanine/methylated guanosine were observed in 20L:4D. These results suggest that a photoperiod would result in no considerable impact on initial meat quality, but extended photoperiods might negatively impact oxidative stability through an alteration of the muscle metabolites.

This study evaluated the effect of dry-aging on quality, palatability, and flavor-related compounds of pork loins. Ten pork loins were obtained at 7 days postmortem, divided into three equal portions, randomly assigned into three different aging methods (wet-aging (W), conventional dry-aging (DA), and UV-light dry-aging (UDA)), and aged for 21 days at 2 °C, 70% RH, and 0.8 m/s airflow. The results showed similar instrumental tenderness values across all treatments (p > 0.05), while DA and UDA had a greater water-holding capacity than WA (p < 0.05). Both DA and UDA were observed to have comparable color stability to WA up to 5 days of retail display (p > 0.05). Greater lipid oxidation was measured in both DA and UDA at the end of display compared to WA (p < 0.05). The UV light minimized microorganisms concentration on both surface and lean portions of UDA compared to other treatments (p < 0.05). The consumer panel was not able to differentiate any sensory traits and overall likeness between the treatments (p > 0.05). Metabolomics analysis, however, identified more flavor-related compounds in dry-aged meat. These findings suggested that dry-aging can be used for pork loins for value-seeking consumers, as it has a potential to generate unique dry-aged flavor in meat with no adverse impacts on meat quality and microbiological attributes.

Gut microbial O-demethylation has been reported for plant-derived dietary compounds containing O-methylated aromatic(s). However, the significance of gut microbial O-demethylation in drug metabolism and disposition remains unexplored. This study examined 64 clinically used oral drugs containing one or more methoxylated aromatics for gut microbial O-demethylation using high-resolution mass spectrometry (HRMS). For 35 of the tested drugs, including the anticancer agent etoposide, we detected metabolites corresponding to O-demethylation (i.e., a mass difference of -14 and its multiples) when individual drugs were incubated with mouse cecal contents. We confirmed that the O-demethylated metabolite (M1) of the model drug etoposide is etoposide catechol using HRMS and proton nuclear magnetic resonance spectroscopy. By testing an in-house collection of 56 gut bacteria individually, we identified seven previously unknown gut bacterial species that exhibit etoposide O-demethylating activity. Etoposide anticancer therapy has been associated with an increased risk of acute myeloid leukemia. We demonstrated that M1 is more genotoxic to myeloid cells when it is orally administered to mice, whereas M1 is less cytotoxic against MCF-7 and HeLa cancer cells than the parent etoposide, suggesting that the gut microbiota may contribute to the secondary genotoxicity of etoposide via O-demethylation. Comparative pharmacokinetic analysis of orally administered etoposide in control and antibiotic-treated mice showed that systemic exposure to etoposide increased 1.9-fold, while M1 exposure decreased 3.7-fold in antibiotic-treated mice, suggesting that gut microbial O-demethylation is a significant determinant of etoposide metabolism and disposition. Collectively, our study reveals the prevalence of gut bacteria with O-demethylation activity, illustrates the contribution of gut microbial O-demethylation to altering drug efficacy and toxicity with the model drug etoposide, and provides a knowledge basis for in-depth characterization of other drugs identified as being susceptible to gut microbial O-demethylation.