Simultaneous Determination of Alternaria Toxins, Ergot Alkaloid Epimers, and Other Major Mycotoxins in Various Food Matrixes by LC-MS/MS.

BACKGROUND: Various food commodities are vulnerable to different types of fungal pathogens and could be contaminated with differential classes of mycotoxins as a result. It is ideal to implement a generic method for the simultaneous determination of multi-mycotoxins in different food matrixes or agricultural products. OBJECTIVE: In this study, a simplified sample preparation procedure and a reliable LC-MS/MS analytical method were developed for the comprehensive measurement of 37 regulated and emerging mycotoxins including five Alternaria toxins (ATs) and six major ergot alkaloids (EAs) and their corresponding epimers. Four different food commodities (baby wheat cereal, peanut, tomato puree, and blended flour) were chosen for method validation to demonstrate the applicability of this analytical method across a wide range of food types. METHODS: Sample extraction was performed using a formic acid-acidified acetonitrile-water (4 + 1, v/v) solution followed by extract dry-down and reconstitution in a water-methanol (1 + 1, v/v) solution for analysis on a biphenyl LC column. Chromatographic analysis was performed using regular acidic LC conditions for baseline separation of ergot alkaloid epimers and completed with a short 11 min cycle time. RESULTS: Accurate quantification was achieved using matrix-matched calibration standards in the range of 0.4 to 400 µg/kg. The recoveries of all mycotoxins (except citrinin) in fortified samples were from 70 to 120%, and the RSD was less than 20%. CONCLUSION: The established workflow was simple and fast for multi-mycotoxin determination in a wide variety of food commodities with LOQs needed to meet the regulatory levels. HIGHLIGHTS: The developed method provided the unique benefit of simultaneous analysis of Alternaria toxins (ATs) and ergot alkaloids (EAs) together with other major regulated mycotoxins.

Exploiting targeted and untargeted approaches for the analysis of bacterial metabolites under altered growth conditions.

In the host, pathogenic microorganisms have developed stress responses to cope with constantly changing environments. Stress responses are directly related to changes in several metabolomic pathways, which could hamper microorganisms' unequivocal identification. We evaluated the effect of various in vitro stress conditions (acidic, basic, oxidative, ethanolic, and saline conditions) on the metabolism of Staphylococcus aureus, Bacillus cereus, and Pseudomonas aeruginosa, which are common lung pathogens. The metabolite profiles of the bacteria were analyzed using liquid chromatography coupled to triple quadrupole and quadrupole time-of-flight mass spectrometry. The advantages of targeted and untargeted analysis combined with univariate and multivariate statistical analysis (principal component analysis, hierarchical cluster analysis, partial least square discriminant analysis, random forest) were combined to unequivocally identify bacterial species. In normal in vitro conditions, the targeted methodology, based on the analysis of primary metabolites, enabled the rapid and efficient discrimination of the three bacteria. In changing in vitro conditions and specifically in presence of the various stressors, the untargeted methodology proved to be more valuable for the global and accurate differentiation of the three bacteria, also considering the type of stress environment within each species. In addition, species-specific metabolites (i.e., fatty acids, polysaccharides, peptides, and nucleotide bases derivatives) were putatively identified. Good intra-day repeatability and inter-day repeatability (< 10% RSD and < 15% RSD, respectively) were obtained for the targeted and the untargeted methods. This untargeted approach highlights its importance in unusual (and less known) bacterial growth environments, being a powerful tool for infectious disease diagnosis, where the accurate classification of microorganisms is sought.

On-line sample preparation for multiclass vitamin, hormone, and mycotoxin determination in chicken egg yolk using LC-MS/MS.

Hen egg yolk from chicken eggs were examined for their mycotoxin, hormone, and fat-soluble vitamin content. A method was developed for multi-class analysis of egg yolk using a dilute/precipitate, centrifuge, and shoot process coupled with on-line sample clean-up using restricted access media with LC-MS/MS. Matrix effects were evaluated and a standard addition protocol with internal standards was chosen for analyte quantitation. With some minor exceptions, the method displayed good linearity (R2 > 0.99), with appropriate limits of detection (0.05-10 ng/g) and limits of quantitation (0.15-30 ng/g) for the analytes tested. Overall, it was discovered there was some variation between the different types of eggs tested in these experiments, especially with regard to their fat-soluble vitamin contents. Low-level mycotoxins were detected in most of the eggs tested. The challenges associated with developing a multi-class compound determination from a single analytical run were elucidated.

Characterization of ethoxylated alcohols in friction reducers using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

RATIONALE: Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) provides detailed information for the analysis of ethoxylated alcohols and polymers. In this study, five friction reducers used in commercial hydraulic fracturing processes were analyzed in their as-received form to identify their ethoxylated alcohol content. The friction reducers were then subjected to lab-simulated downhole conditions. Characterization of friction reducers before and after being subjected to reactive conditions can provide fingerprints associated with produced oilfield waste for source apportionment and information on the stability of these key hydraulic fracturing additives. METHODS: Five different industrially used friction reducers were analyzed for their ethoxylated alcohol content using MALDI-TOF-MS. Three different matrices were assessed for optimal response: α-cyano-4-hydroxycinnamic acid, 2,5-dihydroxybenzoic acid, and 2,5-dihydroxybenzoic acid with 2,2,2-trifluoroethanol (2,5-DHB + E). Reaction times, temperatures, and sample matrices (deionized water, produced water inorganic, produced water, and produced water + shale core) were varied to assess changes in molecular weight distribution and polydispersity of the ethoxylated alcohols relative to their as-received content. RESULTS: A preference for the 2,5-DHB + E matrix was observed. The friction reducers were found to contain ethoxylated alcohols with carbon chain lengths of 12 and 14 with degrees of ethoxylation ranging from 6 to 18. Upon being subjected to 100°C for 24 hours, the ethoxylated alcohols tended to polymerize further, returning higher average molecular weights. Less polymerization was seen in more complex matrices, as supported by dispersity calculations. CONCLUSIONS: Ethoxylated alcohol content was effectively determined in friction reducers using MALDI-TOF-MS. Although this is not a new technique to characterize ethoxylated alcohols, it has proven to be a quick and effective way to determine ethoxylated alcohol content in friction reducers in complex oilfield matrices. This technique can be used as a rapid and straightforward way to determine ethoxylated alcohol content in friction reducers and hydraulic fracturing wastewater for fingerprinting.