Development and Validation of a LC-MS/MS Method for the Determination of Nitrofuran Metabolites in Soft-Shell Turtle Powder Health Food Supplement.

Soft-shell turtle (SST; freshwater terrapin or tortoise) is a popular and important health functional food (HFF) product in many Asian countries. HFFs containing SST must be safe, but several HFFs have been found to be contaminated with dangerous substances, such as nitrofuran metabolites (NFMs). This finding suggests that the consumption of HFFs results in the regular exposure of vulnerable individuals to hazardous substances. Importantly, nitrofuran antibiotics have been banned for use in food-producing animals since the 1990s by the European Union. Thus, in this study, we propose a reliable and quick method to reduce the time required for the detection of four NFMs in SST powder that conventional methods are unable to quantify. Our method involves the derivatization and hydrolysis of SST powder and was validated in accordance with the requirements of European Commission Decision 2002/657/EC. The method achieves an apparent mean recovery of 82.2-108.1%, repeatability of 1.5-3.8%, and reproducibility of 2.2-4.8% for 0.5-10.0 µg kg-1 of 1-aminohydantoin, semicarbazide, 3-amino-2-oxazolidinone, and 3-amino-5-morpholinomethyl-2-oxazolidinone. In addition, linearity was achieved with correlation coefficients of 0.999, and the detection capability (CCß) and decision limit (CCα) were found to be reliable, indicating that this is a fast and accurate method for the analysis of SST powder. The validated method was successfully applied to detect NFMs in SST powder in commercial HHFs.

A simplified modification to rapidly determine the residues of nitrofurans and their metabolites in aquatic animals by HPLC triple quadrupole mass spectrometry.

A simplified method is described for reducing the analysis time of nitrofurans (NFs) and nitrofuran metabolites (NFMs) in the aquatic animals. Most existing HPLC-MS/MS methods are intended only for NFMs and are based on their fast metabolic transformations. We optimized a method for simultaneously detecting major NFs and their metabolites, including nitrovin (NV) that imply use of an optimized buffer solution. The novel method was validated by six different aquatic animal matrices (loach, catfish, shrimp, lobster, scallop, and eel) spiked with the analytes at 0.5, 1.0, and 2.0 µg kg-1. Recovery rates and %RSDs (relative standard deviations) of 82-97% and 1-8% were observed for NFMs, respectively, with values of 70-96% and 1-8% obtained for furazolidone, furaltadone, nitrofurazone, nitrofurantoin, and NV, respectively. Linearity was observed in the 0.1-20 µg L-1 range, with correlation coefficients greater than 0.99 recorded for all compounds. The developed method is sensitive, accurate, easier to use, and faster than previous methods when applied to real samples. To the best of our knowledge, this is the first method that can simultaneously determine NFs and their metabolites, as well as NV, using a single-step extraction process.

Characterization of the antioxidant and anti-inflammatory properties of a polysaccharide-based bioflocculant from Bacillus subtilis F9.

Microbial flocculants are versatile class of novel biomacromolecules with numerous potential industrial applications. This study sought to investigate the antimicrobial, antioxidant, and anti-inflammatory potential of a polysaccharide-based bioflocculant (PBB) extracted from Bacillus subtilis F9. To achieve this, the antioxidant activity of different PBB concentrations(100 µg/mL ̶ 1000 µg/mL) was first examined in vitro using 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl radicals, and superoxide radical scavenging assays. Further, the anti-inflammatory activity of PBB against lipopolysaccharide (LPS; 1 µg/mL)-induced inflammatory mediators released from headkidney (HK)-derived macrophages of Labeo rohita was investigated. Our results revealed that the capacities of 800 µg/mL of PBB to scavenge DPPH, hydroxyl radicals, and superoxide radicals were 81.46 ±â€¯1.37%, 66.34 ±â€¯2.63%, and 78.03 ±â€¯2.46%, respectively, which were slightly higher that observed following treatment with 400 µg/mL of the positive control (ascorbic acid). Further, the radical scavenging capacity of PBB was found to steadily increase with increasing concentrations of PBB. Pre-treatment with PBB also inhibited nitric oxide production in a dose-dependent manner. We next examined the effect of PBB on proinflammatory cytokines (TNF-α, and IL-1ß) and anti-inflammatory cytokines (IL-10, TGF-ß) via qRT-PCR and ELISA. We found that PBB markedly inhibited the LPS-induced mRNA and protein expression levels of TNF-α and IL-1ß, while it significantly increased those of IL-10 and TGF-ß. Further, PBB exhibited an antibacterial activity against multiple food-borne pathogens with minimal inhibitory concentration values in the range of 3 ̶ 11 mg/mL. Importantly, PBB exhibited negligible cytotoxic effects against HK macrophages. Taken together these results suggest that PBB may serve as a natural antioxidant for application in functional therapies and may also be exploited for its anti-inflammatory potential.